Kinase inhibitors useful for the treatment of myleoprolific diseases and other proliferative diseases

ABSTRACT

Compounds of the present invention find utility in the treatment of mammalian cancers and especially human cancers including but not limited to malignant, melanomas, glioblastomas, ovarian cancer, pancreatic cancer, prostate cancer, lung cancers, breast cancers, kidney cancers, cervical carcinomas, metastasis of primary tumor sites, myeloproliferative diseases, leukemias, papillary thyroid carcinoma, non small cell lung cancer, mesothelioma, hypereosinophilic syndrome, gastrointestinal stromal tumors, colonic cancers, ocular diseases characterized by hyperproliferation leading to blindness including various retinopathies, rheumatoid arthritis, asthma, chronic obstructive pulmonary disease, mastocyctosis, mast cell leukemia, a disease caused by c-Abl kinase, oncogenic forms thereof, aberrant fusion proteins thereof and polymorphs thereof, or a disease caused by c-Kit kinase, oncogenic forms thereof, aberrant fusion proteins thereof and polymorphs thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of Provisional Application60/913,216 filed Apr. 20, 2007. This provisional application isincorporated by reference herein in its entirety.

SEQUENCE LISTING

This application contains a Sequence Listing in both paper and computerreadable format in accordance with 37 C.F.R. 1.821 (c) and (e), thecontents of which are hereby incorporated by reference in theirentirety.

FIELD OF THE INVENTION

The present invention relates to novel kinase inhibitors and modulatorcompounds useful for the treatment of various diseases. Moreparticularly, the invention is concerned with such compounds,kinase/compound adducts, methods of treating diseases, and methods ofsynthesis of the compounds. Preferably, the compounds are useful for themodulation of kinase activity of C-Abl, c-Kit, VEGFR, PDGFR kinases,Flt-3, c-Met, FGFR, the HER family and disease polymorphs thereof.

BACKGROUND OF THE INVENTION

Several members of the protein kinase family have been clearlyimplicated in the pathogenesis of various proliferative andmyleoproliferative diseases and thus represent important targets fortreatment of these diseases. Some of the proliferative diseases relevantto this invention include cancer, rheumatoid arthritis, atherosclerosis,and retinopathies. Important examples of kinases which have been shownto cause or contribute to the pathogensis of these diseases includeC-Abl kinase and the oncogenic fusion protein bcr-Abl kinase; c-Kitkinase, PDGF receptor kinase; VEGF receptor kinases; and Flt-3 kinase.

C-Abl kinase is an important non-receptor tyrosine kinase involved incell signal transduction. This ubiquitously expressed kinase—uponactivation by upstream signaling factors including growth factors,oxidative stress, integrin stimulation, and ionizing radiation—localizesto the cell plasma membrane, the cell nucleus, and other cellularcompartments including the actin cytoskeleton (Van Etten, Trends CellBiol. (1999) 9: 179). There are two normal isoforms of Abl kinase:Abl-1A and Abl-1B. The N-terminal half of c-Abl kinase is important forautoinhibition of the kinase domain catalytic activity (Pluk et al, Cell(2002) 108: 247). Details of the mechanistic aspects of thisautoinhibition have recently been disclosed (Nagar et al, Cell (2003)112: 859). The N-terminal myristolyl amino acid residue of Abl-1B hasbeen shown to intramolecularly occupy a hydrophobic pocket formed fromalpha-helices in the C-lobe of the kinase domain. Such intramolecularbinding induces a novel binding area for intramolecular docking of theSH2 domain and the SH3 domain onto the kinase domain, thereby distortingand inhibiting the catalytic activity of the kinase. Thus, an intricateintramolecular negative regulation of the kinase activity is broughtabout by these N-terminal regions of c-Abl kinase. An aberrantdysregulated form of c-Abl is formed from a chromosomal translocationevent, referred to as the Philadelphia chromosome (P. C. Nowell et al,Science (1960) 132: 1497; J. D. Rowley, Nature (1973) 243: 290). Thisabnormal chromosomal translocation leads aberrant gene fusion betweenthe Abl kinase gene and the breakpoint cluster region (BCR) gene, thusencoding an aberrant protein called bcr-Abl (G. Q. Daley et al, Science(1990) 247: 824; M. L. Gishizky et al, Proc. Natl. Acad. Sci. USA (1993)90: 3755; S. Li et al, J. Exp. Med. (1999) 189: 1399). The bcr-Ablfusion protein does not include the regulatory myristolylation site (B.Nagar et al, Cell (2003) 112: 859) and as a result functions as anoncoprotein which causes chronic myeloid leukemia (CML). CML is amalignancy of pluripotent hematopoietic stem cells. The p210 form ofbcr-Abl is seen in 95% of patients with CML, and in 20% of patients withacute lymphocytic leukemia and is exemplified by sequences such as e14a2and e13a2. The corresponding p190 form, exemplified by the sequence e1a2has also been identified. A p185 form has also been disclosed and hasbeen linked to being causative of up to 10% of patients with acutelymphocytic leukemia. It will be appreciated by one skilled in the artthat “p210 form”, “p190 form” and “p185 form” each describe a closelyrelated group of fusion proteins, and that Sequence ID's used herein aremerely representative of each form and are not meant to restrict thescope solely to those sequences.

C-KIT (Kit, CD117, stem cell factor receptor) is a 145 kDa transmembranetyrosine kinase protein that acts as a type-III receptor (Pereira et al.J Carcin. (2005), 4: 19). The c-KIT proto-oncocgene, located onchromosome 4q11-21, encodes the c-KIT receptor, whose ligand is the stemcell factor (SCF, steel factor, kit ligand, mast cell growth factor,Morstyn G, et al. Oncology (1994) 51(2):205. Yarden Y, et al. Embo J(1987) 6(11):3341). The receptor has tyrosine-protein kinase activityand binding of the ligands leads to the autophosphorylation of KIT andits association with substrates such as phosphatidylinositol 3-kinase(Pi3K). Tyrosine phosphorylation by protein tyrosine kinases is ofparticular importance in cellular signalling and can mediate signals formajor cellular processes, such as proliferation, differentiation,apoptosis, attachment, and migration. Defects in KIT are a cause ofpiebaldism, an autosomal dominant genetic developmental abnormality ofpigmentation characterized by congenital patches of white skin and hairthat lack melanocytes. Gain-of-function mutations of the c-KIT gene andthe expression of phosphorylated KIT are found in most gastrointestinalstromal tumors and mastocytosis. Further, almost all gonadalseminomas/dysgerminomas exhibit KIT membranous staining, and severalreports have clarified that some (10-25%) have a c-KIT gene mutation(Sakuma, Y. et al. Cancer Sci (2004) 95:9, 716). KIT defects have alsobeen associated with testicular tumors including germ cell tumors (GCT)and testicular germ cell tumors (TGCT).

The role of c-kit expression has been studied in hematologic and solidtumours, such as acute leukemias (Cortes J. et al. Cancer (2003)97(11):2760) and gastrointestinal stromal tumors (GIST, Fletcher C. D.et al. Hum Pathol (2002) 33(5):459). The clinical importance of c-kitexpression in malignant tumors relies on studies with Gleevec® (imatinibmesylate, STI571, Novartis Pharma AG Basel, Switzerland) thatspecifically inhibits tyrosine kinase receptors (Lefevre G. et al. JBiol Chem (2004) 279(30):31769). Moreover, a clinically relevantbreakthrough has been the finding of anti-tumor effects of this compoundin GIST, a group of tumors regarded as being generally resistant toconventional chemotherapy (de Silva C M, Reid R: Pathol Oncol Res (2003)9(1):13-19). GIST most often become Gleevec resistant and molecularlytargeted small therapies that target c-KIT mutations remain elusive.

c-MET is a unique receptor tyrosine kinase (RTK) located on chromosome7p and activated via its natural ligand hepatocyte growth factor. c-METis found mutated in a variety of solid tumors (Ma P. C. et al. CancerMetastasis (2003) 22:309). Mutations in the tyrosine kinase domain areassociated with hereditary papillary renal cell carcinomas (Schmidt L etal. Nat. Genet. (1997)16:68; Schmidt L, et al. Oncogene (1999) 18:2343),whereas mutations in the sema and juxtamembrane domains are often foundin small cell lung cancers (SCLC; Ma P. C. et al. Cancer Res (2003)63:6272). Many activating mutations are also found in breast cancers(Nakopoulou et al. Histopath (2000) 36(4): 313). The panoply of tumortypes for which c-Met mediated growth has been implicated suggests thisis a target ideally suited for modulation by specific c-MET smallmolecule inhibitors.

The TPR-MET oncogene is a transforming variant of the c-MET RTK and wasinitially identified after treatment of a human osteogenic sarcoma cellline transformed by the chemical carcinogenN-methyl-N′-nitro-N-nitrosoguanidine (Park M. et al. Cell (1986)45:895). The TPR-MET fusion oncoprotein is the result of a chromosomaltranslocation, placing the TPR3 locus on chromosome 1 upstream of aportion of the c-MET gene on chromosome 7 encoding only for thecytoplasmic region. Studies suggest that TPR-MET is detectable inexperimental cancers (e.g. Yu J. et al. Cancer (2000) 88:1801).Dimerization of the M, 65,000 TPR-MET oncoprotein through a leucinezipper motif encoded by TPR leads to constitutive activation of thec-MET kinase (Zhen Z. et al. Oncogene (1994) 9:1691). TPR-MET acts toactivated wild-type c-MET RTK and can activate crucial cellular growthpathways, including the Ras pathway (Aklilu F. et al. Am J Physiol(1996) 271:E277) and the phosphatidylinositol 3-kinase (PI3K)/AKTpathway (Ponzetto C. et al. Mol Cell Biol (1993) 13:4600). Conversely,in contrast to c-MET RTK, TPR-MET is ligand independent, lacks the CBLbinding site in the juxtamembrane region in c-MET, and is mainlycytoplasmic. c-Met immunohistochemical expression seems to be associatedwith abnormal β-catenin expression, and provides good prognostic andpredictive factors in breast cancer patients.

The majority of small molecule kinase inhibitors that have been reportedhave been shown to bind in one of three ways. Most of the reportedinhibitors interact with the ATP binding domain of the active site andexert their effects by competing with ATP for occupancy. Otherinhibitors have been shown to bind to a separate hydrophobic region ofthe protein known as the “DFG-in-conformation” pocket wherein such abinding mode by the inhibitor causes the kinase to adopt the “DFG-out”conformation, and still others have been shown to bind to both the ATPdomain and the “DFG-in-conformation” pocket again causing the kinase toadopt the “DGF-out” conformation. Examples specific to inhibitors of Rafkinases can be found in Lowinger et al, Current Pharmaceutical Design(2002) 8; 2269; Dumas, J. et al., Current Opinion in Drug Discovery &Development (2004) 7: 600; Dumas, J. et al, WO 2003068223 A1 (2003);Dumas, J., et al, WO 9932455 A1 (1999), and Wan, P. T. C., et al, Cell(2004) 116: 855.

Physiologically, kinases are regulated by a commonactivation/deactivation mechanism wherein a specific activation loopsequence of the kinase protein binds into a specific pocket on the sameprotein which is referred to as the switch control pocket. Such bindingoccurs when specific amino acid residues of the activation loop aremodified for example by phosphorylation, oxidation, or nitrosylation.The binding of the activation loop into the switch pocket results in aconformational chance of the protein into its active form (Huse, M. andKuriyan, J. Cell (109) 275)

SUMMARY OF THE INVENTION

Compounds of the present invention find utility in the treatment ofmammalian cancers and especially human cancers including but not limitedto malignant, melanomas, glioblastomas, ovarian cancer, pancreaticcancer, prostate cancer, lung cancers, breast cancers, kidney cancers,cervical carcinomas, metastasis of primary tumor sites,myeloproliferative diseases, leukemias, papillary thyroid carcinoma, nonsmall cell lung cancer, mesothelioma, hypereosinophilic syndrome,gastrointestinal stromal tumors, colonic cancers, ocular diseasescharacterized by hyperproliferation leading to blindness includingvarious retinopathies, rheumatoid arthritis, asthma, chronic obstructivepulmonary disorder, a disease caused by c-Abl kinase, oncogenic formsthereof, aberrant fusion proteins thereof and polymorphs thereof, or adisease caused by c-Kit, oncogenic forms thereof, aberrant fusionproteins thereof and polymorphs thereof.

SECTION 1—DESCRIPTION OF THE PREFERRED EMBODIMENTS

The following descriptions refer to various compounds, stereo-,regioisomers and tautomers of such compounds and individual moieties ofthe compounds thereof.

Cycloalkyl refers to monocyclic saturated carbon rings taken fromcyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptanyl andcyclooctanyl;

Aryl refers to monocyclic or fused bicyclic ring systems characterizedby delocalized π electrons (aromaticity) shared among the ring carbonatoms of at least one carbocyclic ring; preferred aryl rings are takenfrom phenyl, naphlthyl, tetrahydronaphthyl, indenyl, and indanyl;

Heteroaryl refers to monocyclic or fused bicyclic ring systemscharacterized by delocalized π electrons (aromaticity) shared among thering carbon or heteroatoms including nitrogen, oxygen, or sulfur of atleast one carbocyclic or heterocyclic ring; heteroaryl rings are takenfrom, but not limited to, pyrrolyl, furyl, thienyl, oxazolyl, thiazolyl,isoxazolyl, isothiazolyl, imidazolyl, pyrazolyl, oxadiazolyl,thiadiazolyl, triazolyl, tetrazolyl, pyridinyl, pyrimidinyl, pyrazinyl,pyridazinyl, triazinyl, indolyl, indolinyl, isoindolyl, isoindolinyl,indazolyl, benzofuranyl, benzothienyl, benzothiazolyl, benzothiazolonyl,benzoxazolyl, benzoxazolonyl, benzisoxazolyl, benzisothiazolyl,benzimidazolyl, benzimidazolonyl, benztriazolyl, imidazopyridinyl,pyrazolopyridinyl, imidazolonopyridinyl, thiazolopyridinyl,thiazolonopyridinyl, oxazolopyridinyl, oxazolonopyridinyl,isoxazolopyridinyl, isothiazolopyridinyl, triazolopyridinyl,imidazopyrimidinyl, pyrazolopyrimidinyl, imidazolonopyrimidinyl,thiazolopyridiminyl, thiazolonopyrimidinyl, oxazolopyridiminyl,oxazolonopyrimidinyl, isoxazolopyrimidinyl, isothiazolopyrimidinyl,triazolopyrimidinyl, dihydropurinonyl, pyrrolopyrimidinyl, purinyl,pyrazolopyrimidinyl, phthalimidyl, phthalimidinyl, pyrazinylpyridinyl,pyridinopyrimidinyl, pyrimidinopyrimidinyl, cinnolinyl, quinoxalinyl,quinazolinyl, quinolinyl, isoquinolinyl, phthalazinyl, benzodioxyl,benzisothiazoline-1,1,3-trionyl, dihydroquiniolinyl,tetrahydroquinolinyl, dihydroisoquinolyl, tetrahydroisoquinolinyl,benzoazepinyl, benzodiazepinyl, benzoxapinyl, and benzoxazepinyl;

Heterocyclyl refers to monocyclic rings containing carbon andheteroatoms taken from oxygen, nitrogen, or sulfur and wherein there isnot delocalized π electrons (aromaticity) shared among the ring carbonor heteroatoms; heterocyclyl rings include, but are not limited to,oxetanyl, azetadinyl, tetrahydrofuranyl, pyrrolidinyl, oxazolinyl,oxazolidinyl, thiazolinyl, thiazolidinyl, pyranyl, thiopyranyl,tetrahydropyranyl, dioxalinyl, piperidinyl, morpholinyl,thiomorpholinyl, thiomorpholinyl S-oxide, thiomorpholinyl S-dioxide,piperazinyl, azepinyl, oxepinyl, diazepinyl, tropanyl, and homotropanyl;

Poly-aryl refers to two or more monocyclic or fused aryl bicyclic ringsystems characterized by delocalized π electrons (aromaticity) sharedamong the ring carbon atoms of at least one carbocyclic ring wherein therings contained therein are optionally linked together;

Poly-heteroaryl refers to two or more monocyclic or fused bicyclicsystems characterized by delocalized π electrons (aromaticity) sharedamong the ring carbon or heteroatoms including nitrogen, oxygen, orsulfur of at least one carbocyclic or heterocyclic ring wherein therings contained therein are optionally linked together, wherein at leastone of the monocyclic or fused bicyclic rings of the poly-heteroarylsystem is taken from heteroaryl as defined broadly above and the otherrings are taken from either aryl, heteroaryl, or heterocyclyl as definedbroadly above;

Poly-heterocyclyl refers to two or more monocyclic or fused bicyclicring systems containing carbon and heteroatoms taken from oxygen,nitrogen, or sulfur and wherein there is not delocalized π electrons(aromaticity) shared among the ring carbon or heteroatoms wherein therings contained therein are optionally linked, wherein at least one ofthe monocyclic or fused bicyclic rings of the poly-heteroaryl system istaken from heterocyclyl as defined broadly above and the other rings aretaken from either aryl, heteroaryl, or heterocyclyl as defined broadlyabove;

Alkyl refers to straight or branched chain C1-C6alkyls;

Halogen refers to fluorine, chlorine, bromine, and iodine;

Alkoxy refers to —O-(alkyl) wherein alkyl is defined as above;

Alkoxylalkyl refers to -(alkyl)-O-(alkyl) wherein alkyl is defined asabove;

Alkoxylcarbonyl refers to —C(O)O-(alkyl) wherein alkyl is defined asabove;

CarboxylC1-C6alkyl refers to —C(O)-alkyl wherein alkyl is defined asabove;

Substituted in connection with a moiety refers to the fact that afurther substituent may be attached to the moiety to any acceptablelocation on the moiety.

The term salts embraces pharmaceutically acceptable salts commonly usedto form alkali metal salts of free acids and to form addition salts offree bases. The nature of the salt is not critical, provided that it ispharmaceutically-acceptable. Suitable pharmaceutically-acceptable acidaddition salts may be prepared from an inorganic acid or from an organicacid. Examples of such inorganic acids are hydrochloric, hydrobromic,hydroiodic, nitric, carbonic, sulfuric and phosphoric acid. Appropriateorganic acids may be selected from aliphatic, cycloaliphatic, aromatic,arylaliphatic, and heterocyclyl containing carboxylic acids and sulfonicacids, examples of which are formic, acetic, propionic, succinic,glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic,glucuronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic,anthranilic, mesylic, stearic, salicylic, p-hydroxybenzoic,phenylacetic, mandelic, embonic (pamoic), methanesulfonic,ethanesulfonic, benzenesulfonic, pantothenic, toluenesulfonic,2-hydroxyethanesulfonic, sulfanilic, cyclohexylaminosulfonic, algenic,3-hydroxybutyric, galactaric and galacturonic acid. Suitablepharmaceutically-acceptable salts of free acid-containing compounds ofFormula I include metallic salts and organic salts. More preferredmetallic salts include, but are not limited to appropriate alkali metal(group Ia) salts, alkaline earth metal (group IIa) salts and otherphysiological acceptable metals. Such salts can be made from aluminum,calcium, lithium, magnesium, potassium, sodium and zinc. Preferredorganic salts can be made from primary amines, secondary amines,tertiary amines and quaternary ammonium salts, including in part,tromethamine, diethylamine, tetra-N-methylammonium,N,N′-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine,ethylenediamine, meglumine(N-methylglucamine) and procaine.

The term prodrug refers to derivatives of active compounds which revertin vivo into the active form. For example, a carboxylic acid form of anactive drug may be esterified to create a prodrug, and the ester issubsequently converted in vivo to revert to the carboxylic acid form.See Ettmayer et. al, J. Med. Chem (2004) 47: 2393 and Lorenzi et. al, J.Pharm. Exp. Therapeutic (2005) 883 for reviews.

Structural, chemical and stereochemical definitions are broadly takenfrom IUPAC recommendations, and more specifically from Glossary of Termsused in Physical Organic Chemistry (IUPAC Recommendations 1994) assummarized by P. Müller, Pure Appl. Chem., 66, 1077-1184 (1994) andBasic Terminology of Stereochemistry (IUPAC Recommendations 1996) assummarized by G. P. Moss Pure and Applied Chemistry, 68, 2193-2222(1996). Specific definitions are as follows:

Atropisomers are defined as a subclass of conformers which can beisolated as separate chemical species and which arise from restrictedrotation about a single bond.

Regioisomers or structural isomers are defined as isomers involving thesame atoms in different arrangements.

Enatiomers are defined as one of a pair of molecular entities which aremirror images of each other and non-superimposable.

Diastereomers or diastereoisomers are defined as stereoisomers otherthan enantiomers. Diastereomers or diastereoisomers are stereoisomersnot related as mirror images.

Diastereoisomers are characterized by differences in physicalproperties, and by some differences in chemical behavior towards achiralas well as chiral reagents.

Tautomerism is defined as isomerism of the general form

where the isomers (called tautomers) are readily interconvertible; theatoms connecting the groups X,Y,Z are typically any of C, H, O, or S,and G is a group which becomes an electrofuge or nucleofuge duringisomerization. The commonest case, when the electrofuge is H⁺, is alsoknown as “prototropy”.

Tautomers are defined as isomers that arise from tautomerism,independent of whether the isomers are isolable.

1. First Aspect of the Invention—Compounds, Methods, Preparations andAdducts

and wherein the pyridine ring may be optionally substituted with one ormore R20 moieties;

each D is individually taken from the group consisting of C, CH, C—R20,N-Z3, N, O and S, such that the resultant ring is taken from the groupconsisting of triazolyl, isoxazolyl, isothiazolyl, oxazolyl, andthiadiazolyl;

wherein E is selected from the group consisting of phenyl, pyridyl, andpyrimidinyl;

E may be optionally substituted with one or two R16 moieties;

wherein A is a ring system selected from the group consisting of phenyl,naphthyl, cyclopentyl, cyclohexyl, G1, G2, and G3;

G1 is a heteroaryl taken from the group consisting of pyrrolyl, furyl,thienyl, oxazolyl, thiazolyl, isoxazol-4-yl, isoxazol-5-yl,isothiazolyl, imidazolyl, pyrazolyl, oxadiazolyl, thiadiazolyl,triazolyl, tetrazolyl, pyrazinyl, pyridazinyl, triazinyl, pyridinyl, andpyrimidinyl;

G2 is a fused bicyclic heteroaryl taken from the group consisting ofindolyl, indolinyl, isoindolyl, isoindolinyl, indazolyl, benzofuranyl,benzothienyl, benzothiazolyl, benzothiazolonyl, benzoxazolyl,benzoxazolonyl, benzisoxazolyl, benzisothiazolyl, benzimidazolyl,benzimidazolonyl, benztriazolyl, imidazopyridinyl, pyrazolopyridinyl,imidazolonopyridinyl, thiazolopyridinyl, thiazolonopyridinyl,oxazolopyridinyl, oxazolonopyridinyl, isoxazolopyridinyl,isothiazolopyridinyl, triazolopyridinyl, imidazopyrimidinyl,pyrazolopyrimidinyl, imidazolonopyrimidinyl, thiazolopyridiminyl,thiazolonopyrimidinyl, oxazolopyridiminyl, oxazolonopyrimidinyl,isoxazolopyrimidinyl, isothiazolopyrimidinyl, triazolopyrimidinyl,dihydropurinonyl, pyrrolopyrimidinyl, purinyl, pyrazolopyrimidinyl,phthalimidyl, phthalimidinyl, pyrazinylpyridinyl, pyridinopyrimidinyl,pyrimidinopyrimidinyl, cinnolinyl, quinoxalinyl, quinazolinyl,quinolinyl, isoquinolinyl, phthalazinyl, benzodioxyl,benzisothiazoline-1,1,3-trionyl, dihydroquinolinyl,tetrahydroquinolinyl, dihydroisoquinolyl, tetrahydroisoquinolinyl,benzoazepinyl, benzodiazepinyl, benzoxapinyl, and benzoxazepinyl;

G3 is a heterocyclyl taken from the group consisting of oxetanyl,azetadinyl, tetrahydrofuranyl, pyrrolidinyl, oxazolinyl, oxazolidinyl,imidazolonyl, pyranyl, thiopyranyl, tetrahydropyranyl, dioxalinyl,piperidinyl, morpholinyl, thiomorpholinyl, thiomorpholinyl S-oxide,thiomorpholinyl S-dioxide, piperazinyl, azepinyl, oxepinyl, diazepinyl,tropanyl, and homotropanyl;

the A ring may be optionally substituted with one or two R2 moieties;

X is selected from the group consisting of —O—, —S(CH₂)_(n)—,—N(R3)(CH₂)_(n)—, —(CH₂)_(p)—, and wherein the carbon atoms of—(CH₂)_(n)—, —(CH₂)_(p)—, of X may be further substituted by oxo or oneor more C1-C6alkyl moieties;

when A, G1, G2 or G3 has one or more substitutable sp2-hybridized carbonatoms, each respective sp2 hybridized carbon atom may be optionallysubstituted with a Z1 substituent;

when A, G1, C2 or G3 has one or more substitutable sp3-hybridized carbonatoms, each respective sp3 hybridized carbon atom may be optionallysubstituted with a Z2 substituent;

when A, G1, G2 or G3 has one or more substitutable nitrogen atoms, eachrespective nitrogen atom may be optionally substituted with a Z4substituent;

each Z1 is independently and individually selected from the groupconsisting of C1-6alkyl, branched C3-C7alkyl, C3-C8cycloalkyl, halogen,fluoroC1-C6alkyl wherein the alkyl moiety can be partially or fullyfluorinated, cyano, C1-C6alkoxy, fluoroC1-C6alkoxy wherein the alkylmoiety can be partially or fully fluorinated, —(CH₂)_(n)OH, oxo,C1-C6alkoxyC1-C6alkyl, (R4)₂N(CH₂)_(n)—, (R3)₂N(CH₂)_(n)—,(R4)₂N(CH₂)_(q)N(R4)(CH₂)_(n)—, (R4)₂N(CH₂)_(q)O(CH₂)_(n)—, (R3)₂NC(O)—,(R4)₂NC(O)—, (R4)₂NC(O)C1-C6alkyl-, —(R4)NC(O)R8, C1-C6alkoxycarbonyl-,-carboxyC1-C6alkyl, C1-C6alkoxycarbonylC1-C6alkyl-, (R3)₂NSO₂—, —SOR3,(R4)₂NSO₂—, —N(R4)SO₂R8, —O(CH₂)_(q)OC1-C6alkyl, —SO₂R3, —SOR4,—C((O)R8, —C(O)R6, —C(═NOH)R6, —C(═NOR3)R6, —(CH₂)_(n)N(R4)C(O)R8,—N(R3)(CH₂)_(q)O-alkyl, —N(R3)(CH₂)_(q)N(R4)₂, nitro, —CH(OH)CH(OH)R4,—C(═NH)N(R4)₂, —C(═NOR3)N(R4)₂, —NHC(═NH)R8, R17 substituted G3, R17substituted pyrazolyl and R17 substituted imidazolyl;

in the event that Z1 contains an alkyl or alkylene moiety, such moietiesmay be further substituted with one or more C1-C6alkyls;

each Z2 is independently and individually selected from the groupconsisting of aryl, C1-C6alkyl, C3-C8cycloalkyl, branched C3-C7alkyl,hydroxyl, hydroxyC1-C6alkyl-, cyano, (R3)₂N—, (R4)₂N—,(R4)₂NC1-C6alkyl-, (R4)₂NC2-C6alkylN(R4)(CH₂)_(n)—,(R4)₂NC2-C6alkylO(CH₂)_(n)—, (R3)₂NC(O)—, (R4)₂NC(O)—,(R4)₂NC(O)—C1-C6alkyl-, carboxyl, -carboxyC1-C6alkyl,C1-C6alkoxycarbonyl-, C1-C6alkoxycarbonylC1-C6alkyl-, (R3)₂NSO₂—,(R4)₂NSO₂—, —SO₂R8, —(CH₂)_(n)N(R4)C(O)R8, —C(O)R8, ═O, ═NOH, and═N(OR6);

in the event that Z2 contains an alkyl or alkylene moiety, such moietiesmay be further substituted with one or more C1-C6alkyls;

each Z3 is independently and individually selected from the groupconsisting of H, C1-C6alkyl, branched C3-C7alkyl, C3-C8cycloalkyl,fluoroC1-C6alkyl wherein the alkyl moiety can be partially or fullyfluorinated, hydroxyC2-C6alkyl-, C1-C6alkoxycarbonyl-, —C(O)R8,R5C(O)(CH₂)_(n)—, (R4)₂NC(O)—, (R4)₂NC(O)C1-C6alkyl-,R8C(O)N(R4)(CH₂)_(q)—, (R3)₂NSO₂—, (R4)₂NSO₂—, —(CH₂)_(q)N(R3)₂, and—(CH₂)_(q)N(R4)₂;

each Z4 is independently and individually selected from the groupconsisting of C1-C6alkyl, branched C3-7alkyl, hydroxyC2-C6alkyl-,C1-C6alkoxyC2-C6alkyl-, (R4)₂N—C2-C6alkyl-,(R4)₂N—C2-C6alkylN(R4)—C2-C6alkyl-,(R4)₂N—C2-C6alkyl-O—C2-C6alkyl-(R4)₂NC(O)C1-C6alkyl-, carboxyC1-C6alkyl,C1-C6alkoxycarbonylC1-C6alkyl-, —C2-C6alkylN(R4)C(O)R8, R8-C(═NR3)—,—SO₂R8, and —COR8;

in the event that Z4 contains an alkyl or alkylene moiety, such moietiesmay be further substituted with one or more C1-C6alkyls;

each R2 is selected from the group consisting of H, C1-C6alkyl, branchedC3-C8alkyl, R19 substituted C3-C8cycloalkyl-, fluoroC1-C6alkyl- whereinthe alkyl is fully or partially fluorinated, halogen, cyano,C1-C6alkoxy-, and fluoroC1-C6alkoxy- wherein the alkyl group is fully orpartially fluorinated, hydroxyl substituted C1-C6alkyl-, hydroxylsubstituted branched C3-C8alkyl-, cyano substituted C1-C6alkyl-, cyanosubstituted branched C3-C8alkyl-, (R3)₂NC(O)C1-C6alkyl-, and(R3)₂NC(O)C3-C8 branched alkyl-;

wherein each R3 is independently and individually selected from thegroup consisting of H, C1-C6alkyl, branched C3-C7alkyl, andC3-C8cycloalkyl;

each R4 is independently and individually selected from the groupconsisting of H, C1-C6alkyl, hydroxyC1-C6alkyl-, dihydroxyC1-C6alkyl-,C1-C6alkoxyC1-C6alkyl-, branched C3-C7alkyl, branchedhydroxyC1-C6alkyl-, branched C1-C6alkoxyC1-C6alkyl-, brancheddihydroxyC1-C6alkyl-, —(CH₂)_(p)N(R7)₂, —(CH₂)_(p)C(O)N(R7)₂,—(CH₂)_(n)C(O)OR3, and R19 substituted C3-C8cycloalkyl-;

each R5 is independently and individually selected from the groupconsisting of

and wherein the symbol (##) is the point of attachment to Z3;

each R6 is independently and individually selected from the groupconsisting of C1-C6alkyl, branched C3-C7alkyl, and R19 substitutedC3-C8cycloalkyl-;

each R7 is independently and individually selected from the groupconsisting of H, C1-C6alkyl, hydroxyC2-C6alkyl-, dihydroxyC2-C6alkyl-,C1-C6alkoxyC2-C6alkyl-, branched C3-C7alkyl, branchedhydroxyC2-C6alkyl-, branched C1-C6alkoxyC2-C6alkyl-, brancheddihydroxyC2-C6alkyl-, —(CH₂)_(n)C(O)OR3, R19 substitutedC3-C8cycloalkyl- and —(CH₂)_(n)R17;

each R8 is independently and individually selected from the groupconsisting of C1-C6alkyl, branched C3-C7alkyl, fluoroC1-C6alkyl- whereinthe alkyl moiety is partially or fully fluorinated, R19 substitutedC3-C8cycloalkyl-, —OH, C1-C6alkoxy, —N(R3)₂, and —N(R4)₂;

each R10 is independently and individually selected from the croupconsisting of —CO₂H, —CO₂C1-C6alkyl, —C(O)N(R4)₂, OH, C1-C6alkoxy, and—N(R4)₂;

each R16 is independently and individually selected from the groupconsisting of H, C1-C6alkyl, branched C3-C7alkyl, R19 substitutedC3-C8cycloalkyl-, halogen, fluoroC1-C6alkyl- wherein the alkyl moietycan be partially or fully fluorinated, cyano, hydroxyl, C1-C6alkoxy,fluoroC1-C6alkoxy- wherein the alkyl moiety can be partially or fullyfluorinated, —N(R3)₂, —N(R4)₂, R3 substituted C2-C3alkynyl- and nitro;

each R17 is independently and individually selected from the groupconsisting of H, C1-C6alkyl, branched C3-C7alkyl, R19 substitutedC3-C8cycloalkyl-, halogen, fluoroC1-C6alkyl- wherein the alkyl moietycan be partially or Filly fluorinated, cyano, hydroxyl, C1-C6alkoxy,fluoroC1-C6alkoxy- wherein the alkyl moiety can be partially or fullyfluorinated, —N(R3)₂, —N(R4)₂, and nitro;

each R19 is independently and individually selected from the groupconsisting of H, OH and C1-C6alkyl;

each R20 is independently and individually selected from the groupconsisting of C1-C6alkyl, branched C3-C7alkyl, R19 substitutedC3-C8cycloalkyl-, halogen, fluoroC1-C6alkyl- wherein the alkyl moietycan be partially or fully fluorinated, cyano, hydroxyl, C1-C6alkoxy,fluoroC1-C6alkoxy- wherein the alkyl moiety can be partially or fullyfluorinated, —N(R3)₂, —N(R4)₂, —N(R3)C(O)R3, —C(O)N(R3)₂ and nitro andwherein two R4 moieties independently and individually taken from thegroup consisting of C1-C6alkyl, branched C3-C6alkyl, hydroxyalkyl-, andalkoxyalkyl and attached to the same nitrogen heteroatom may cyclize toform a C3-C7 heterocyclyl ring;

k is 0 or 1; n is 0-6; p is 1-4; q is 2-6; r is 0 or 1; t is 1-3; v is 1or 2; m is 0-2;

and stereo-, regioisomers and tautomers of such compounds.

1.1 Compounds of Formula Ia which Exemplify Referred D Moieties

In a preferred embodiment of compounds of formula Ia, said compoundshave preferred

moieties of the formula:

wherein the symbol (**) indicates the point of attachment to thepyridine ring.

1.1.1 Compounds of Formula Ia which Exemplify Preferred A Moieties

In a preferred embodiment of compounds of formula Ia, said compoundshave structures of formula Ib

wherein A is any possible isomer of pyrazole.

1.1.2 Compounds of Formula Ia which Exemplify Preferred A and R16Moieties

In a more preferred embodiment of compounds of formula Ib, saidcompounds have structures of formula Ic

1.1.3 Compounds of Formula Ia which Exemplify Preferred A and R16Moieties

In a more preferred embodiment of compounds of formula Ib, saidcompounds have structures of formula Id

1.1.4 Compounds of Formula Ia which Exemplify Preferred A and R16Moieties

In a more preferred embodiment of compounds of formula Ib, saidcompounds have structures of formula Ie

1.1.5 Compounds of Formula Ia which Exemplify Preferred A Moieties

In a more preferred embodiment of compounds of formula Ia, saidcompounds have structures of formula If

1.1.6 Compounds of Formula Ia which Exemplify Preferred A and R16Moieties

In a more preferred embodiment of compounds of formula Ia, saidcompounds have structures of formula Ig

1.1.7 Compounds of Formula Ia which Exemplify Preferred A Moieties

In a preferred embodiment of compounds of formula Ia, said compoundshave structures of formula Ih

wherein A is selected from the group consisting of any possible isomerof phenyl and pyridine.

1.1.8 Compounds of Formula Ia which Exemplify Preferred A and R16Moieties

In a more preferred embodiment of compounds of formula Ih, saidcompounds have structures of formula Ii

1.1.9 Compounds of Formula Ia which Exemplify Preferred A and R16Moieties

In a more preferred embodiment of compounds of formula Ih, saidcompounds have structures of formula Ij

1.1.10 Compounds of Formula Ia which Exemplify Preferred A Moieties

In a more preferred embodiment of compounds of formula Ia, saidcompounds have structures of formula Ik

1.1.11 Compounds of Formula Ia which Exemplify Preferred A and R16Moieties

In a more preferred embodiment of compounds of formula Ik, saidcompounds have structures of formula Il

1.1.12 Most Preferred Compounds of Formula Ia

1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(oxazol-5-yl)pyridin-4-yloxy)phenyl)urea,1-(4-(2-(1H-1,2,3-triazol-4-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)urea,1-(4-(2-(1H-1,2,4-triazol-1-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(3-tert-butylisoxazol-5-yl)ureaand1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(oxazol-2-yl)pyridin-4-yloxy)phenyl)urea.

1.2 Methods

1.2a Methods of Protein Modulation

The invention includes methods of modulating kinase activity of avariety of kinases, e.g. C-Abl kinase, bcr-Abl Kinase, Flt-3, VEGFR-2kinase mutants, c-Met, c-Kit, PDGFR and the HER family of kinases. Thekinases may be wildtype kinases, oncogenic forms thereof, aberrantfusion proteins thereof or polymorphs of any of the foregoing. Themethod comprises the step of contacting the kinase species withcompounds of the invention and especially those set forth in sectionssection 1. The kinase species may be activated or unactivated, and thespecies may be modulated by phosphorylations, sulfation, fatty acidacylations glycosylations, nitrosylation, cystinylation (i.e. proximalcysteine residues in the kinase react with each other to form adisulfide bond) or oxidation. The kinase activity may be selected fromthe group consisting of catalysis of phospho transfer reactions,inhibition of phosphorylation, oxidation or nitrosylation of said kinaseby another enzyme, enhancement of dephosphorylation, reduction ordenitrosylation of said kinase by another enzyme, kinase cellularlocalization, and recruitment of other proteins into signaling complexesthrough modulation of kinase conformation.

1.2b Treatment Methods

The methods of the invention also include treating individuals sufferingfrom a condition selected from the group consisting of cancer andhyperproliferative diseases. These methods comprise administering tosuch individuals compounds of the invention, and especially those ofsection 1, said diseases including, but not limited to, malignantmelanomas, glioblastomas, ovarian cancer, pancreatic cancer, prostatecancer, lung cancers, breast cancers, kidney cancers, cervicalcarcinomas, metastasis of primary tumor secondary sites,myeloproliferative diseases, leukemias, papillary thyroid carcinoma, nonsmall cell lung cancer, mesothelioma, hypereosinophilic syndrome,gastrointestinal stromal tumors, colonic cancers, ocular diseasescharacterized by hyperproliferation leading to blindness includingvarious retinopathies including diabetic retinopathy and age-relatedmacular degeneration, rheumatoid arthritis, asthma, chronic obstructivepulmonary disorder, mastocytosis, mast cell leukemia, a disease causedby c-Abl kinase, oncogenic forms thereof, aberrant fusion proteinsthereof and polymorphs thereof, or a disease caused by a c-Kit kinase,oncogenic forms thereof, aberrant fusion proteins thereof and polymorphsthereof. The administration method is not critical, and may be from thegroup consisting of oral, parenteral, inhalation, and subcutaneous.

1.3 Pharmaceutical Preparations

The compounds of the invention, especially those of section 1 may form apart of a pharmaceutical composition by combining one or more suchcompounds with a pharamaceutically acceptable carrier. Additionally, thecompositions may include an additive selected from the group consistingof adjuvants, excipients, diluents, and stabilizers.

SECTION 2. SYNTHESIS OF COMPOUNDS OF THE PRESENT INVENTION

The compounds of the invention are available by the procedures andteachings of WO 2006/071940, incorporated by reference, and by thegeneral synthetic methods illustrated in the Schemes below and theaccompanying examples.

As indicated in Scheme 1, ureas of general formula 1 can be readilyprepared by the union of amines of general formula 2 with isocyanates 3or isocyanate surrogates, for example trichloroethyl carbamates (4) orisopropenyl carbamates (5). Preferred conditions for the preparation ofcompounds of general formula 1 involve heating a solution of 4 or 5 with2 in the presence of a tertiary base such as diisopropylethylamine,triethylamine or N-methylpyrrolidine in a solvent such asdimethylformamide, dimethylsulfoxide, tetrahydrofuran or 1,4-dioxane ata temperature between 50 and 100° C. for a period of time ranging from 1hour to 2 days.

As shown in Scheme 2, isocyanates 3 can be prepared from amines A-NH₂ 6with phosgene, or a phosgene equivalent such as diphosgene, triphosgene,or N,N-dicarbonylimidazole. Trichloroethyl carbamates 4 and isopropenylcarbamates 5 are readily prepared from amines A-NH₂, (6) by acylationwith trichloroethyl chloroformate or isopropenyl chloroformate bystandard conditions familiar to those skilled in the art. Preferredconditions for the preparation of 4 and 5 include include treatment ofcompound 6 with the appropriate chloroformate in the presence ofpyridine in an aprotic solvent such as dichloromethane or in thepresence of aqueous hydroxide or carbonate in a biphasic aqueous/ethylacetate solvent system.

Additionally, compounds of formula 1 can also be prepared fromcarboxylic acids 7 by the intermediacy of in-situ generated acyl azides(Curtius rearrangement) as indicated in Scheme 3. Preferred conditionsfor Scheme 3 include the mixing of acid 7 with amine 2 anddiphenylphosphoryl azide in a solvent such as 1,4-dioxane ordimethylformamide in the presence of base, such as triethylamine andraising the temperature of the reaction to about 80-120° C. to affectthe Curtius rearrangement.

By analogy to Schemes 1 and 3 above, it will be recognized by thoseskilled in the art that the compounds of formula 1 can also be preparedby the union of amines A-NH₂ 6 with isocyanates 8 (Scheme 4).Isocyanates 8 can be prepared from general amines 2 by standardsynthetic methods. Suitable methods for example, include reaction of 2with phosgene, or a phosgene equivalent such as diphosgene, triphosgene,or N,N-dicarbonylimidazole. In addition to the methods above forconverting amines 2 into isocynates 8, the isocyanates 8 can also beprepared in situ by the Curtius rearrangement and variants thereof.Those skilled in the art will further recognize that isocycanates 8 neednot be isolated, but may be simply generated in situ. Accordingly, acid9 can be converted to compounds of formula 1 either with or withoutisolation of 8. Preferred conditions for the direct conversion of acid 9to compounds of formula 1 involve the mixing of acid 9, amine A-NH₂ 6,diphenylphosphoryl azide and a suitable base, for example triethylamine,in an aprotic solvent, for example dioxane. Heating said mixture to atemperature of between 80 and 120° C. provides the compounds of formula1.

Additionally, compounds of formula 1 can also be prepared from amines 2by first preparing stable isocyanate equivalents, such as carbamates(Scheme 5). Especially preferred carbamates include trichloroethylcarbamates (10) and isopropenyl carbamates (11) which are readilyprepared from amine 2 by reaction with trichloroethyl chloroformate orisopropenyl chloroformate respectively using standard conditionsfamiliar to those skilled in the art. Further reaction of carbamates 10or 11 with amine A-NH₂ 6 provides compounds of formula 1. Those skilledin the art will further recognize that certain carbamates can also beprepared from acid 9 by Curtius rearrangement and trapping with analcoholic co-solvent. For example, treatment of acid 9 (Scheme 5) withdiphenylphosphoryl azide and trichloroethanol at elevated temperatureprovides trichloroethyl carbamate 10.

Many methods exist for the preparation of amines A-NH₂ 6 and acidsA-CO₂H 7, depending on the nature of the A-moiety. Indeed, many suchamines (6) and acids (7) useful for the preparation of compounds offormula 1 are available from commercial vendors. Some non-limitingpreferred synthetic methods for the preparation of amines 6 and acids 7are outlined in the following schemes and accompanying examples.

As illustrated in Scheme 6, Z4-substituted pyrazol-5-yl amines 14 (apreferred aspect of A-NH₂ 6, Scheme 2) are available by the condensationof hydrazines 12 and beta-keto nitrites 13 in the presence of a strongacid. Preferred conditions for this transformation are by heating inethanolic HCl. Many such hydrazines 12 are commercially available.Others can be prepared by conditions familiar to those skilled in theart, for example by the diazotization of amines followed by reductionor, alternately from the reduction of hydrazones prepared from carbonylprecursors.

Another preferred method for constructing Z4-substituted pyrazoles isillustrated by the general preparation of pyrazole acids 19 and 20.(Scheme 7), aspects of of general acid A-CO₂H 7 (Scheme 3). As indicatedin Scheme 7, pyrazole 5-carboxylic esters 17 and 18 can be prepared bythe alkylation of pyrazole ester 16 with Z4-X 15, wherein X represents aleaving group on a Z4 moiety such as a halide, triflate, or othersulfonate. Preferred conditions for the alkylation of pyrazole 16include the use of strong bases such as sodium hydride, potassiumtert-butoxide and the like in polar aprotic solovents such asdimethylsulfoxide, dimethylformamide or tetrahydrofuran. Z4-substitutedpyrazoles 17 and 18 are isomers of one another and can both be preparedin the same reactions vessel and separated by purification methodsfamiliar to those skilled in the art. The esters 17 and 18 in turn canbe converted to acids 19 and 20 using conditions familiar to thoseskilled in the art, for example saponification in the case of ethylesters, hydrogenation in the case of benzyl esters or acidic hydrolysisin the case of tert-butyl esters.

Scheme 8 illustrates the preparation of pyrazole amine 25, a furtherexample of general amine A-NH₂ 6. Acid-catalyzed condensation ofR2-substituted hydrazine 21 with 1,1,3,3-tetramethoxypropane 22 providesR2-substituted pyrazole 23. Those skilled in the art will furtherrecognize that R2-substituted pyrazole 23 can also be prepared by directalkylation of pyrazole. Pyrazole 23 can be regioselectively nitrated toprovide nitro-pyrazole 24 by standard conditions familiar to thoseskilled in the art. Finally, hydrogenation of nitro-pyrazole 24employing a hydrogenation catalyst, such as palladium or nickel providespyrazole amine 25, an example of general amine A-NH₂ 6.

Additional pyrazoles useful for the synthesis of compounds of formula 1can be prepared as described in Scheme 9. Thus, keto-ester 26 can bereacted with N,N-dimethylformamide dimethyl acetal to provide 27.Reaction of 27 with either 21 or 28 (wherein P is an acid-labileprotecting group) in the presence of acid provides 29 or 30. Inpractice, both 29 and 30 can be obtained from the same reaction and canbe separated by standard chromatographic conditions. In turn, esters 29and 30 can be converted to acids 31 and 32 respectively as previouslydescribed in Scheme 7.

In a manner similar to Scheme 9, NH-pyrazole 34 can be prepared byreaction of acrylate 33 with hydrazine (Scheme 10). Alkylation of 34with R2-X 35 as described above for Scheme 7 provides mixtures ofpyrazole esters 36 and 37 which are separable by standardchromatographic techniques. Further conversion of esters 36 and 37 toacids 38 and 39 can be accomplished as described above in Scheme 7.

General amines 6 containing an isoxazole ring can be prepared asdescribed in Scheme 11. Thus, by analogy to Scheme 6, reaction ofketo-nitrile 9 with hydroxylamine can provide both the 5-aminoisoxazole40 and 3-aminoisoxazole 41. Preferred conditions for the formation of5-aminoisoxazole 40 include the treatment of 9 with hydroxylamine in thepresence of aqueous sodium hydroxide, optionally in the presence of analcoholic co-solvent at a temperature between 15 and 100° C. Preferredconditions for the formation of 3-aminoisoxazole 41 include thetreatment of 9 with hydroxylamine hydrochloride in a polar solvent suchas water, an alcohol, dioxane or a mixture thereof at a temperaturebetween 15 and 100° C.

Amines 2 useful for the invention can be synthesized according tomethods commonly known to those skilled in the art. Amines of generalformula 2 contain three rings and can be prepared by the stepwise unionof three monocyclic subunits as illustrated in the followingnon-limiting Schemes. Scheme 12 illustrates one mode of assembly inwhich an E-containing subunit 42 is combined with the central pyridinering 43 to provide the bicyclic intermediate 44. In one aspect thisgeneral Scheme, the “M” moiety of 42 represents a hydrogen atom of aheteroatom on the X linker that participates in a nucleophilic aromaticsubstitution reaction with monocycle 43. Such reactions may befacilitated by the presence of bases (for example, potassiumtert-butoxide), thus M may also represent a suitable counterion (forexample potassium, sodium, lithium, or cesium) within an alkoxide,sulfide or amide moiety. Alternately, the “M” group can represent ametallic species (for example, copper, boron, tin, zirconium, aluminum,magnesium, lithium, silicon, etc.) on a carbon atom of the X2 moietythat can undergo a transition-metal-mediated coupling with monocycle 43.

The “Y” group of monocyclic species 42 is an amine or an aminesurrogate, such as an amine masked by a protecting group (“P” in formula45), a nitro group, or a carboxy acid or ester that can be used toprepare an amine via known rearrangement. Examples of suitableprotecting groups “P” include but are not limited to tert-butoxycarbonyl(Boc), benzyloxycarbonyl (Cbz), and acetamide. In the instances whereinthe “Y”-group of intermediate 42 is not an amine, the products of Scheme11 will be amine surrogates such as 45 or 46 that can be converted toamine 2 by a deprotection, reduction or rearrangement (for example,Curtius rearrangement) familiar to those skilled in the all.

In these instances, the “LG” of monocycle 43 represents a moiety thatcan either be directly displaced in a nucleophilic substitution reaction(with or without additional activation) or can participate in atransition-mediated union with fragment 42. The W group of monocycle 43or bicycle 44 represents a moiety that allows the attachment of thepyrazole. In one aspect, the “W” group represents a halogen atom thatwill participate in a transition-metal-mediated coupling with apre-formed heterocyclic reagent (for example a boronic acid or ester, orheteroaryl stannane) to give rise to amine 2. In another aspect, the “W”group of 43 and 44 represents a functional group that can be convertedto a five-membered heterocycle by an annulation reaction. Non-limitingexamples of such processes would include the conversion of a cyano,formyl, carboxy, acetyl, or alkynyl moiety into a pyrazole moiety. Itwill be understood by those skilled in the art that such annulations mayin fact be reaction sequences and that the reaction arrows in Scheme 11may represent either a single reaction or a reaction sequence.Additionally, the “W” group of 44 may represent a leaving group (halogenor triflate) that can be displaced by a nucleophilic nitrogen atom of apyrazole ring.

Some non-limiting examples of general Scheme 12 are illustrated in theSchemes below. Scheme 13 illustrates the preparation of pyrazole 51, anexample of general amine 2. In Scheme 13, commercially available3-fluoro-4-aminophenol (47) is reacted with potassium tert-butoxide and2,4-dichloropyridine 48 to provide chloropyridine 49. The preferredsolvent for this transformation is dimethylacetamide at a temperaturebetween 80 and 100° C. Subsequent union of chloropyridine 49 with thecommercially available oxazole-4-boronic acid pinacol ester 50 in thepresence of a palladium catalyst, preferablytetrakis(triphenylphosphine)palladium, provides oxazole amine 51.

Scheme 14 illustrates a non-limiting example of Scheme 12 wherein the“W” group is a leaving group for nucleophilic aromatic substitution.Thus, amine 53, an example of general amine 2, can be prepared fromgeneral intermediate 49 by reaction with 1,2,4-triazole (52). Preferredconditions include the use of polar aprotic solvents such as1-methyl-2-pyrrolidinone, dimethylacetamide, or dimethylsulfoxide in thepresence of non-nucleophilic bases such as potassium carbonate, sodiumhydride, 1,8-diaza-bicyclo[5.4.0]undec-7-ene (DBU), and the like.Preferred temperatures are from ambient temperature up to about 250° C.and may optionally include the use of microwave irradiation orsonication. Those skilled in the art will recognize that the generalmethods of scheme 14 can be used to prepare additional triazole isomersby employing either 1,2,4-triazole 52, or alternatively, by employing1,2,3-triazole in place of 52.

Scheme 15 illustrates the preparation of amine 55 and 56, non-limitingexamples of general amine of formula 2, by way of an annulation sequenceaccording to general Scheme 12. Conversion of chloropyridine 49 intoalkyne 53 can be accomplished by Sonogashira cross-coupling withtrimethylsilylacetylene, followed by aqueous hydrolysis of thetrimethylsilyl group, conditions familiar to those skilled in the art.Further reaction of alkyne 53 with azidomethyl pivalate (54) in thepresence of copper sulfate and sodium ascorbate provides theN-pivaloylymethyl triazole amine 55. (see Loren, et. al. Synlett,(2005), 2847). Deprotection under standard conditions, preferably diluteaqueous sodium hydroxide, provides 56. Alternatively, the amine 55 canbe used directly to produce ureas of formula 1 prior to the removal ofthe N-pivaloylmethyl protecting group.

Additional examples of general amines of formula 2 can be prepared asillustrated in Scheme 16. Thus, by analogy to Scheme 12, the generalintermediate 40 can be converted by palladium-mediated Stille-couplinginto oxazoles 57 or 59 by reaction with the tributylstannanes 58 (see:Cheng et al., Biorg. Med. Chem. Lett., 2006, 2076) or 60 (AldrichChemical). Preferred palladium catalysts for the Stille reactionsinclude dichlorobis(triplhenylphosphine)palladium,dichloro[11′-bis(diphenylphosphino)ferrocene]palladium andtetrakis(triphenylphosphine)palladium. Similarly, isoxazoles 61 and 63can be obtained by the palladium-catalyzed reaction of 40 with4-isoxazoleboronic acid pinacol ester 62 (Frontier Scientific) ortributylstannane 64 (see: Sakamoto, et al. Tetrahedron, 1991, 5111).

As an extension of Schemes 12, 13 and 16, amines of general formula 2containing an isothiazole ring can also be prepared by the methodsdescribed above. Scheme 17 shows a non-limiting example wherein apalladium-catalyzed Stille reaction of trimethylstannane 65 (see:Wentland, et al. J. Med. Chem., 1993, 1580) with 40 can provideisothiazole 67. In a similar fashion, palladium-catalyzed Suzuki-crosscoupling between 40 and the boronate ester 66 (see: Blackaby, et al.,U.S. Pat. No. 7,030,128) gives rise to isothiazole amine 68.

Additional preferred synthetic methods for the preparation of compoundsof formula 1 are found in the following examples.

SECTION 3. EXAMPLES

General Method A: To a stirring solution of the carboxylic acid (0.24mmol) and TEA (1.2 mmol) in 1,4-dioxane (4.5 mL) at RT was added DPPA(0.29 mmol). After stirring for 0.5 h at RT, the appropriate amine (0.71mmol) was added and the reaction was stirred with heating at 100° C. for2 h. The reaction was cooled to RT, diluted with brine (15 mL) andextracted with EtOAc (3×30 mL). The combined organic layers were dried(MgSO₄) and concentrated. The residue was purified by chromatography toafford the target compound.

General Method B: To a solution of the starting pyrazole amine (1 eq) inEtOAc were added 2,2,2-trichloroethylchloroformate (1.1 eq) andsaturated NaHCO₃ (2-3 eq) at 0° C. After stirring for 3 h at RT, thelayers were separated and the aqueous layer extracted with EtOAc. Thecombined organic extracts were washed with brine, dried (Na₂SO₄) andconcentrated under vacuum to yield the crude TROC carbamate of thepyrazole amine.

To the TROC carbamate (1 eq) in DMSO were added diisopropylethylamine (2eq), the appropriate amine (2 eq) and the mixture was stirred at 60° C.for 16 h or until all the starting carbamate was consumed. Water wasadded to the mixture and the product was extracted with EtOAc (2×25 mL).The combined organic extracts were washed with brine solution, dried(Na₂SO₄) and concentrated to yield crude product, which was purified bycolumn chromatography to yield the target compound.

Example A1

A suspension of 3-fluoro-4-aminophenol (8.0 g, 63.0 mmol) indimethylacetamide (80 mL) was de-gassed in vacuo and treated withpotassium tert-butoxide (7.3 g, 65 mmol). The resultant mixture wasstirred at RT for 30 min. 2,4-Dichloropyridine (8 g, 54 mmol) was addedand the mixture was heated to 80° C. for 12 h. The solvent was removedunder reduced pressure to give a residue which was partitioned betweenwater and EtOAc (3×100 mL). The organic layers were washed withsaturated brine, dried (MgSO₄), concentrated in vacuo and purified bysilica gel column chromatography to give4-(2-chloro-pyridin-4-yloxy)-2-fluoro-phenylamine (11 g, 86% yield). ¹HNMR (300 MHz, DMSO-d6), δ 8.24 (d, J=5.7 Hz, 1 H), 7.00 (dd, J=9.0, 2.7Hz, 1 H), 6.89-6.73 (m, 4 H), 5.21 (br s, 2 H); MS (ESI) m/z: 239.2(M+H+).

To a degassed solution of4-(2-chloropyridin-4-yloxy)-2-fluorobenzenamine (0.801 g, 3.36 mmol) inDMF (2 mL) and TEA (2 mL) was added ethynyltrimethylsilane (0.929 ml,6.71 mmol), trans-dichloro-bis(triphenyl phosphine)palladium(0) (0.236g, 0.336 mmol) and copper (I) iodide (0.064 g, 0.336 mmol) and themixture was stirred at 90° C. for 16 h. Water (60 ml) was added to themixture, product was extracted with EtOAc (2×45 ml) and the combinedorganics were washed with brine, dried (Na₂SO₄) concentrated to affordcrude product. The product was dissolved in methanol (10 ml), K₂CO₃ (0.5g) was added and the mixture was stirred at RT for 2 h. Solvent wasremoved, to the crude residue was added water (60 mL) and EtOAc (40 ml),the layers were separated and the aqueous layer was extracted with EtOAc(1×30 mL). The combined organic layer was washed with brine, dried(Na₂SO₄) and concentrated to afford crude product which was purified bycolumn chromatography (ethylacetate/hexane) to afford4-(2-ethynylpyridin-4-yloxy)-2-fluorobenzenamine as a thick residue(0.56 g, 73% yield). ¹H NMR (400 MHz, DMSO-d₆): δ 8.37 (d, J=6.0 Hz,1H), 6.98 (dd, J=8.0 Hz, 2.4 Hz, 1H), 6.95 (d, J=6.0 Hz, 1H), 6.87 (dd,J=6.0 Hz, 2.4 Hz, 1H), 6.81-6.73 (m, 2H), 5.20 (brs, 2H), 4.03 (s, 1H);MS (ESI) m/z: 229.1 (M+H⁺).

Sodium azide (1.942 g, 29.9 mmol) was added to a suspension ofchloromethyl pivalate (3.00 g, 19.92 mmol) in water (5 mL) and stirredvigorously at 90° C. for 16 h. The reaction mixture was diluted withwater (20 mL) and EtOAc (20 ml). The organic layer was washed withbrine, dried (Na₂SO₄) and concentrated to afford azidomethyl pivalate asa liquid (2 g, 64% yield). ¹H NMR (400 MHz, Acetone-d₆): δ 5.23 (s, 2H),1.22 (s, 9H).

To a suspension of azidomethyl pivalate (0.075 g, 0.477 mmol),4-(2-ethynylpyridin-4-yloxy)-2-fluorobenzenamine (0.109 g, 0.477 mmol)in t-butanol (0.6 mL) and water (0.6 mL) was added sodium ascorbate(0.021 g, 0.095 mmol). Copper(II)sulfate in water (0.048 ml, 0.048 mmol)was added to the above suspension and the dark red mixture was stirredfor 3 h at RT. It was diluted with water (30 mL) and EtOAc (20 mL), thelayers were separated and the aqueous layer was extracted with EtOAc(2×15 mL). The combined organics were washed with brine, dried (Na₂SO₄)and concentrated to afford(4-(4-(4-amino-3-fluorophenoxy)pyridin-2-yl)-1H-1,2,3-triazol-1-yl)methylpivalate as a red solid. (0.165 g, 90% yield). ¹H NMR (400 MHz,DMSO-d₆): δ 8.54 (s, 1H), 8.46 (brs, 1H), 7.60 (s, 1H), 6.98 (d, J=8.8Hz, 1H), 6.94 (d, J=3.6 Hz, 1H), 6.83-6.81 (m, 2H), 6.42 (s, 2H), 4.78(s, 2H), 1.17 (s, 9H); MS (ESI) m/z: 386.1 (M+H⁺).

Example A2

To a solution of 4-(2-chloropyridin-4-yloxy)-2-fluorobenzenamine fromExample A1 (1.0 g, 4.2 mmol) in NMP (10 ml) was added DBU (0.94 mL, 6.3mmol) and 1,2,4-triazol sodium salt (0.57 g, 6.3 mmol) and the mixturewas heated overnight under argon atmosphere at 160° C. The reactionmixture cooled to RT, diluted with water (100 mL) and the solution wasextracted with EtOAc (3×). The organics were combined and washed withLiCl solution and brine (2×), dried (Na₂SO₄) and concentrated in vacuo.The residue was slurried in EtOAc (5 mL), the solid was filtered andwashed with EtOAc to obtain a mixture of product and SM. The filtratewas concentrated in vacuo, the residue was slurried in CH₂Cl₂, filteredand washed with CH₂Cl₂ to obtain4-(2-(1H-1,2,4-triazol-1-yl)pyridin-4-yloxy)-2-fluorobenzenamine (0.35g). MS (ESI) m/z: 272.2 (M+H⁺).

Example A3

4-(2-Chloropyridin-4-yloxy)-2-fluorobenzenamine from Example A1 (150 mg,0.629 mmol), 2-(tri-n-butylstannyl)oxazole (0.132 ml, 0.629 mmol) andPdCl₂(dppf)-CH₂Cl₂ (51.3 mg, 0.063 mmol) were combined in DMF (3 ml)under Ar and stirred with heating at 90° C. After 3 h, the completedreaction was cooled to RT and treated with satd. aq. KF (5 ml; preparedfrom equal portions of KF.H₂O and H₂O) and stirred at RT for 1 h. Thesuspension was diluted with EtOAc and filtered through Celite®, rinsingforward with EtOAc. The filtrate was diluted with H₂O and the layerswere separated. The aqueous was extracted with EtOAc (2×). The combinedorganics were washed with brine (2×), dried (MgSO₄), concentrated invacuo and purified by flash column chromatography (EtOAc/hexanes) toafford 80 mg of 2-fluoro-4-(2-(oxazol-2-yl)pyridin-4-yloxy)benzenamine(0.295 mmol, 47% yield) as an oil that solidified on standing. ¹H NMR(400 MHz, DMSO-d₆): δ 8.52 (d, J=5.8 Hz, 1H), 8.26 (s, 1H), 7.40 (d,J=2.3 Hz, 1H), 7.38 (s, 1H), 7.05-7.02 (m, 2H), 6.88-6.78 (m, 2H), 5.23(s, 2H); ); MS (ESI) m/z: 272.1 (M+H⁺).

Example A4

In a 3:1 mix of DMF:H20 (6 mL) was placed4-(2-chloropyridin-4-yloxy)-2-fluorobenzenamine from example A1 (245 mg,1.026 mmol), cesium carbonate (1.337 g, 4.10 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)oxazole (300 mg, 1.538mmol) and tetrakistriphenylphosphine Pd(0) (178 mg, 0.154 mmol). The mixwas degassed, placed under Ar, warmed to 80° C. and stirred overnight.The reaction was cooled to RT, diluted with water (20 mL) and extractedwith ethyl acetate (3×20 mL). The combined organic phases were washedwith brine, dried (Na₂SO₄) and concentrated in vacuo to give2-fluoro-4-(2-(oxazol-5-yl)pyridin-4-yloxy)benzenamine (415 mg, 149%yield) as a dark oil. LC and LCMS shows mostly desired product as wellas triphenylphosphine oxide as a by product. Used as is. MS (ESI) m/z:272.1 (M+H⁺).

Example A5

In a microwave reaction vial,4-(2-ethynylpyridin-4-yloxy)-2-fluorobenzenamine from Example A1 (0.201g, 0.881 mmol) was dissolved in THF (4 mL). Acetaldoxime (0.078 g, 1.321mmol), triethylamine (0.246 ml, 1.761 mmol), and1-chloropyrrolidine-2,5-dione (0.176 g, 1.321 mmol) were added and themixture was stirred at 130° C. for 45 min under microwave irradiation.An additional 1.5 eq each of acetaldoxime and1-chloropyrrolidine-2,5-dione were added and microwave heating washeated for an additional 45 min at 130° C. This process repeated onemore time. The mixture was poured into a biphasic solution of water (40mL) and EtOAc (30 mL). The organic layer was separated and the aqueouslayer was extracted with EtOAc (2×20 mL). The combined organics werewashed with brine, dried (Na₂SO₄) and concentrated. The residue waspurified by silica gel chromatography to afford2-fluoro-4-(2-(3-methylisoxazol-5-yl)pyridin-4-yloxy)benzenamine (58 mg,23% yield) as light red colored residue. MS (ESI) m/z: 286.1 (M+H⁺).

Example B1

To an aqueous solution of sodium hydroxide solution (40.00 g, 1 mol, in200 ml of water) was added hydroxylamine hydrochloride (24.00 g, 346mmol) and pivaloylacetonitrile (40.00 g, 320 mmol). The resultingsolution was stirred at 50° C. for 3 hrs. The reaction mixture cooledand the resultant white crystalline solid filtered, washed with waterand dried to provide 3-t-butylisoxazol-5-amine as a white crystallinesolid (34 g, yield 76% yield). ¹H NMR (DMSO-d₆) δ 6.41 (brs, 2H), 4.85(s, 1H), 1.18(s, 9H): LC-MS (ES, m/z, M+H) 141.3.

Example 1

Using General Method A, 3-tert-butyl-1-methyl-1H-pyrazole-5-carboxylicacid (0.054 g, 0.3 mmol), Example A1 (0.1 g, 0.25 mmol), triethylamine(0.76 g, 0.75 mmol) and DPPA (0.137 g, 0.5 mmol) were combined andpurified by column chromatography (ethylacetate/hexanes) to afford(4-(4-(4-(3-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)ureido)-3-fluorophenoxy)pyridin-2-yl)-1H-1,2,3-triazol-1-yl)methylpivalate as a white solid (0.115 g, 82% yield). ¹H NMR (400 MHz,DMSO-d₆): δ 8.89 (s, 1H), 8.84 (s, 1H), 8.68 (s, 1H), 8.50 (d, J=5.6 Hz,1H), 8.19 (t, J=8.8 Hz, 1H), 7.46 (d, J=2.4 Hz, 1H), 7.33 (dd, J=12.0Hz, 2.8 Hz, 1H), 7.06 (dd, J=8.8 Hz, 1.6 Hz, 1H), 6.97 (dd, J=5.6 Hz,2.4 Hz, 1H), 6.35 (s, 2H), 6.07 (s, 1H), 3.60 (s, 3H), 1.19 (s, 9H),1.10 (s, 9H); MS (ESI) m/z: 565.2 (M+H⁺).

To a solution of(4-(4-(4-(3-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)ureido)-3-fluorophenoxy)pyridin-2-yl)-1H-1,2,3-triazol-1-yl)methylpivalate (0.11 g, 0.195 mmol) in MeOH (1 mL) was added 2M NaOH (0.4 mL)and the mixture was stirred for 30 min at RT. Solvent was removed, cruderesidue was diluted with water (5 mL) and neutralized with 50% aqueousacetic acid (1 ml). The resultant solid was filtered and dried to afford1-(4-(2-(1H-1,2,3-triazol-4-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)ureaas a white solid (75 mg, 85% yield). ¹H NMR (400 MHz, DMSO-d₆): δ 8.89(s, 1H), 8.84 (s, 1H), 8.68 (s, 1H), 8.49 (d, J=6.0 Hz, 1H), 8.36 (brs,1H), 8.19 (t, J=9.2 Hz, 1H), 7.42 (s, 1H), 7.33 (dd, J=11.6 Hz, 2.4 Hz,1H), 7.06-7.04 (m, 1H), 6.94 (dd, J=5.6 Hz, 2.0 Hz, 1H), 6.07 (s, 1H),3.60 (s, 3H), 1.19 (s, 9H); MS (ESI) m/z: 451.1 (M+H⁺).

Example 2

To a solution of 2,2,2-trichloroethyl 3-tert-butylisoxazol-5-ylcarbamate(0.080 g, 0.25 mmol), formed via General method B from Example B1, indioxane (3 mL) was added Example A2 (70 mg, 0.25 mmol) and1-methylpyrrolidine (22 mg, 0.25 mmol). The reaction mixture was heatedovernight at 65° C. The reaction mixture cooled to RT, concentrated invacuo, DCM (2 mL) was added and the slurry was stirred for 1 hour. Thesolid was filtered and air dried to obtain1-(4-(2-(1H-1,2,4-triazol-1-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(3-tert-butylisoxazol-5-yl)urea.¹H NMR (400 MHz, DMSO-d₆): δ 10.3 (s, 1H), 9.56 (s, 1H), 8.70 (s, 1H),8.34 (s, 1H), 8.29 (d, J=6.0 Hz, 1H), 8.03 (t, J=9.2 Hz, 1H), 7.68 (dd,J=2.0, and 5.6 Hz, 1H), 7.57 (d, J=1.2 Hz, 1H), 7.26 (dd, J=2.8, and12.0 Hz, 1H), 7.03 (m, 1H), 6.05 (s, 1H), 1.24 (s, 9H); MS (ESI) m/z:438.1 (M+H⁺).

Example 3

To a solution of 3-(t-butyl)-1-methyl-1H-pyrazole-5-carboxylic acid(0.054 g, 0.295 mmol) in dioxane (3 ml) was added TEA (0.123 ml, 0.885mmol) followed by DPPA (0.095 ml, 0.442 mmol). The mixture was stirredat RT for 30 min and then treated with a solution of Example A3 (0.080g, 0.295 mmol) in dioxane (3.00 ml). The reaction was then placed in anoil bath preheated to 100° C. and stirred with heating overnight. Thecompleted reaction was cooled to RT. Without aqueous workup, thereaction mixture was purified directly by reverse phase chromatography(MeCN (w/0.1% TFA)/H₂O (w/0.1% TFA)) to afford1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(oxazol-2-yl)pyridin-4-yloxy)phenyl)ureaof 96.8% purity. MS (ESI) m/z: 451.1 (M+H⁺).

Example 4

Using General Method A, 3-tert-butyl-1-methyl-1H-pyrazole-5-carboxylicacid (205 mg, 1.127 mmol), triethylamine (415 mg, 4.10 mmol), Example A4[max theoretical yield from previous reaction] (278 mg, 1.025 mmol) andDPPA (338 mg, 1.23 mmol) were combined and purified by reverse phasechromatography (acetonitrile/water/−0.1% TFA) to give a residue whichwas treated with 10% potassium carbonate (10 mL) and extracted withethyl acetate (3×25 mL). The combined organic phases were washed withbrine (25 mL), dried (Na2SO4) and concentrated in vacuo to give1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(oxazol-5-yl)pyridin-4-yloxy)phenyl)urea(131 mg, 28% yield) as a film. ¹H NMR (400 MHz, DMSO-d₆): δ 1.15 (s, 9H), 3.60 (s, 3 H), 6.07 (s, 1 H), 6.91-6.93 (m, 1 H), 7.04-7.07 (m, 1H), 7.23 (d, 1 H), 7.30-7.34 (m, 1 H), 7.78 (s, 1 H), 8.18 (t, 1 H),8.48-8.51 (m, 2 H), 8.85 (d, 1 H), 8.90 (s, 1 H); MS (ESI) m/z: 451.1(M+H⁺).

Example 5

Using general method A, 3-tert-butyl-1-methyl-1H-pyrazole-5-carboxylicacid (0.042 g, 0.23 mmol), Example A5 (0.51 g, 0.18 mmol), triethylamine(0.54 g, 0.53 mmol) and DPPA (0.1 g, 0.35 mmol) were combined andpurified by column chromatography using (EtOAc/hexanes) to afford1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(3-methylisoxazol-5-yl)pyridin-4-yloxy)phenyl)ureaas white solid (0.015 g, 18% yield). ¹H NMR (400 MHz, DMSO-d₆): δ 8.89(s, 1H), 8.85 (s, 1H), 8.56 (d, J=6.0 Hz, 1H), 8.20 (t, J=8.8 Hz, 1H),7.38 (d, J=2.4 Hz, 1H), 7.34 (dd, J=12.0 Hz, 2.4 Hz, 1H), 7.08-7.05 (m,1H), 7.03-7.00 (m, 1H), 6.97 (s, 1H), 6.07 (s, 1H), 3.60 (s, 3H), 2.82(s, 3H), 1.19 (s, 9H); MS (ESI) m/z: 465.1 (M+H⁺).

Using the synthetic procedures and methods described herein and methodsknown to those skilled in the art, the following compounds are made:1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(4-methyloxazol-2-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(3-methyl-1,2,4-thiadiazol-5-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(isoxazol-5-yl)pyridin-4-yloxy)phenyl)urea,1-(4-(2-(4H-1,2,4-triazol-4-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(isothiazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(3-methyl-1H-1,2,4-triazol-1-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl-3-(2-fluoro-4-(2-(isothiazol-3-yl)pyridin-4-yloxy)phenyl)urea1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(isothiazol-5-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(4-methyl-2H-1,2,3-triazol-2-yl)pyridin-4-yloxy)phenyl)urea,1-(5-tert-butylisoxazol-3-yl)-3-(2-fluoro-4-(2-(4-methyl-4H-1,2,4-triazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(5-methyl-1,2,4-thiadiazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(4-methyl-4H-1,2,4-triazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(3-methyl-3H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(isoxazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(isoxazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butylisoxazol-5-yl)-3-(2-fluoro-4-(2-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butylisoxazol-5-yl)-3-(2-fluoro-4-(2-(5-methyl-1,2,4-thiadiazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butylisoxazol-5-yl)-3-(2-fluoro-4-(2-(4-methyl-4H-1,2,4-triazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butylisoxazol-5-yl)-3-(2-fluoro-4-(2-(3-methyl-3H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butylisoxazol-5-yl)-3-(2-fluoro-4-(2-(isoxazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butylisoxazol-5-yl)-3-(2-fluoro-4-(2-(isoxazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(1-tert-butyl-1H-pyrazol-4-yl)-3-(2-fluoro-4-(2-(isoxazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(4-methyloxazol-2-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(3-methyl-1,2,4-thiadiazol-5-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(isoxazol-5-yl)pyridin-4-yloxy)phenyl)urea,1-(4-(2-(4H-1,2,4-triazol-4-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(isothiazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(3-methyl-1H-1,2,4-triazol-1-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(isothiazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(isothiazol-5-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(4-methyl-2H-1,2,3-triazol-2-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(5-methyl-1,2,4-thiadiazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(4-methyl-4H-1,2,4-triazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(3-methyl-3H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(isoxazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(isoxazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butylisoxazol-5-yl)-3-(2-fluoro-4-(2-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butylisoxazol-5-yl)-3-(2-fluoro-4-(2-(5-methyl-1,2,4-thiadiazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butylisoxazol-5-yl)-3-(2-fluoro-4-(2-(4-methyl-4H-1,2,4-triazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butylisoxazol-5-yl)-3-(2-fluoro-4-(2-(3-methyl-3H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butylisoxazol-5-yl)-3-(2-fluoro-4-(2-(isoxazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butylisoxazol-5-yl)-3-(2-fluoro-4-(2-(isoxazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(1-tert-butyl-1H-pyrazol-4-yl)-3-(2-fluoro-4-(2-(isoxazol-4-yl(pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(1-methyl-1H-1,2,4-triazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butylisoxazol-5-yl)-3-(2-fluoro-4-(2-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butylisoxazol-5-yl)-3-(2-fluoro-4-(2-(1-methyl-1H-1,2,4-triazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(1-tert-butyl-1H-pyrazol-4-yl)-3-(2-fluoro-4-(2-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(1-tert-butyl-1H-pyrazol-4-yl)-3-(2-fluoro-4-(2-(1-methyl-1H-1,2,4-triazol-3-yl)pyridin-4-yloxy)phenyl)urea,1-(2-fluoro-4-(2-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)-3-(5-(trifluoromethyl)pyridin-3-yl)urea,1-(2-fluoro-4-(2-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)-3-(5-isopropylpyridin-3-yl)urea,1-(2-fluoro-4-(2-(1-methyl-1H-1,2,4-triazol-3-yl)pyridin-4-yloxy)phenyl)-3-(5-(trifluoromethyl)pyridin-3-yl)urea,1-(2-fluoro-4-(2-(1-methyl-1H-1,2,4-triazol-3-yl)pyridin-4-yloxy)phenyl)-3-(5-isopropylpyridin-3-yl)urea,1-(2-fluoro-4-(2-(3-methyl-1,2,4-thiadiazol-5-yl)pyridin-4-yloxy)phenyl)-3-(5-(trifluoromethyl)pyridin-3-yl)urea,1-(2-fluoro-4-(2-(3-methyl-12,4-thiadiazol-5-yl)pyridin-4-yloxy)phenyl)-3-(5-isopropylpyridin-3-yl)urea,1-(2-fluoro-4-(2-(3-methyl-1H-1,2,4-triazol-1-yl)pyridin-4-yloxy)phenyl)-3-(5-(trifluoromethyl)pyridin-3-yl)urea,1-(2-fluoro-4-(2-(3-methyl-1H-1,2,4-triazol-1-yl)pyridin-4-yloxy)phenyl)-3-(5-isopropylpyridin-3-yl)urea,1-(4-(2-(4H-1,2,4-triazol-4-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(5-isopropylpyridin-3-yl)urea,1-(4-(2-(4H-1,2,4-triazol-4-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(5-(trifluoromethyl)pyridin-3-yl)urea,1-(2-fluoro-4-(2-(3-methyl-1H-1,2,4-triazol-1-yl)pyridin-4-yloxy)phenyl)-3-(5-isopropylpyridin-3-yl)urea,1-(2-fluoro-4-(2-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)-3-(5-(trifluoromethyl)pyridin-3-yl)urea,1-(2-fluoro-4-(2-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)-3-(5-isopropylpyridin-3-yl)urea,1-(2-fluoro-4-(2-(5-methyl-1,2,4-thiadiazol-3-yl)pyridin-4-yloxy)phenyl)-3-(5-(trifluoromethyl)pyridin-3-yl)urea,1-(2-fluoro-4-(2-(5-methyl-1,2,4-thiadiazol-3-yl)pyridin-4-yloxy)phenyl)-3-(5-isopropylpyridin-3-yl)urea,1-(2-fluoro-4-(2-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)-3-(5-(trifluoromethyl)pyridin-3-yl)urea,1-(2-fluoro-4-(2-(isoxazol-4-yl)pyridin-4-yloxy)phenyl)-3-(5-isopropylpyridin-3-yl)urea,1-(2-fluoro-4-(2-(isoxazol-4-yl)pyridin-4-yloxy)phenyl)-3-(5-(trifluoromethyl)pyridin-3-yl)urea,1-(2-fluoro-4-(2-(2-methyl-2H-1,2,3-triazol-4-yl)pyridin-4-yloxy)phenyl)-3-(5-isopropylpyridin-3-yl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-5-(2-(1-methyl-1H-pyrazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(5-(2-(1H-pyrazol-4-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)urea,1-(5-(2-(1H-pyrazol-3-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-5-(2-(oxazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-5-(2-(oxazol-2-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-5-(2-(oxazol-5-yl)pyridin-4-yloxy)phenyl)urea,1-(1-tert-butyl-1H-pyrazol-4-yl)-3-(2-fluoro-5-(2-(3-methylisoxazol-5-yl)pyridin-4-yloxy)phenyl)urea,1-(1-tert-butyl-1H-pyrazol-4-yl)-3-(2-fluoro-5-(2-(isoxazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(1-tert-butyl-1H-pyrazol-4-yl)-3-(2-fluoro-5-(2-(isothiazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(1-tert-butyl-1H-pyrazol-4-yl)-3-(2-fluoro-5-(2-(3-methylisothiazol-5-yl)pyridin-4-yloxy)phenyl)urea,1-(5-(2-(1H-1,2,4-triazol-1-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(1-tert-butyl-1H-pyrazol-4-yl)urea,1-(5-(2-(4H-1,2,4-triazol-3-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(1-tert-butyl-1H-pyrazol-4-yl)urea,1-(5-(2-(1H-1,2,3-triazol-1-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(1-tert-butyl-1H-pyrazol-4-yl)urea,1-(5-(2-(1,3,4-thiadiazol-2-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(1-tert-butyl-1H-pyrazol-4-yl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(oxazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(oxazol-5-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(3-methylisoxazol-5-yl)pyridin-4-yloxy)phenyl)urea,1-(1-tert-butyl-1H-pyrazol-4-yl)-3-(2-fluoro-4-(2-(3-methylisothiazol-5-yl)pyridin-4-yloxy)phenyl)urea,1-(3-tert-butylisoxazol-5-yl)-3-(2-fluoro-4-(2-(1-methyl-1H-imidazol-4-yl)pyridin-4-yloxy)phenyl)urea,1-(4-(2-(1H-1,2,4-triazol-1-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)urea,1-(4-(2-(4H-1,2,4-triazol-3-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(1-tert-butyl-1H-pyrazol-4-yl)urea,1-(4-(2-(1H-1,2,3-triazol-1-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(1-tert-butyl-1H-pyrazol-4-yl)urea,1-(4-(2-(1,3,4-thiadiazol-2-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(1-tert-butyl-1H-pyrazol-4-yl)urea,1-(4-(2-(1H-1,2,4-triazol-1-yl)pyridin-4-yloxy)-2-fluorophenyl)-3-(3-tert-butylisoxazol-5-yl)urea,and1-(3-tert-butyl-1-methyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(oxazol-2-yl)pyridin-4-yloxy)phenyl)urea.

SECTION 4. BIOLOGICAL DATA

Abl Kinase (SEQ ID NO:1) Assay

Activity of Abl kinase (SEQ ID NO:1) was determined by following theproduction of ADP from the kinase reaction through coupling with thepyruvate kinase/lactate dehydrogenase system (e.g., Schindler, et al.Science (2000) 289, 1938-1942). In this assay, the oxidation of NADH(thus the decrease at A_(340nm) was continuously monitoredspectrophometrically. The reaction mixture (100 μl) contained Abl kinase(1 nM. Abl from deCode Genetics), peptide substrate (EAIYAAPFAKKK, 0.2mM), MgCl₂ (10 mM), pyruvate kinase (4 units), lactate dehydrogenase(0.7 units), phosphoenol pyruvate (1 mM), and NADH (0.298 mM) in 90 mMTris buffer containing 0.2% octyl-glucoside and 3.5% DMSO, pH 7.5. Testcompounds were incubated with Abl (SEQ ID NO:1) and other reactionreagents at 30° C. for 2 h before ATP (500 μM) was added to start thereaction. The absorption at 340 nm was monitored continuously for 2hours at 30° C. on Polarstar Optima plate reader (BMG). The reactionrate was calculated using the 1.0 to 2.0 h time frame. Percentinhibition was obtained by comparison of reaction rate with that of acontrol (i.e. with no test compound). IC₅₀ values were calculated from aseries of percent inhibition values determined at a range of inhibitorconcentrations using software routines as implemented in the GraphPadPrism software package.

Abl kinase (SEQ ID NO: 1)GTSMDPSSPNYDKWEMERTDITMKHKLGGGQYGEVYEGVWKKYSLTVAVKTLKEDTMEVEEFLKEAAVMKEIKHPNLVQLLGVCTREPPFYIITEFMTYGNLLDYLRECNRQEVNAVVLLYMATQISSAMEYLEKKNFIHRDLAARNCLVGENHLVKVADFGLSRLMTGDTYTAHAGAKFPIKWTAPESLAYNKFSIKSDVWAFGVLLWEIATYGMSPYPGIDLSQVYELLEKDYRMERPEGCPEKVYELMRACWQWNPSDRPSFAEIHQAFETMFQE

Abl Kinase (SEQ ID NO:2) Assay

Activity of T3 15I Abl kinase (SEQ ID NO:2) was determined by followingthe production of ADP from the kinase reaction through coupling with thepyruvate kinase/lactate dehydrogenase system (e.g., Schindler, et al.Science (2000) 289, 1938-1942). In this assay, the oxidation of NADH(thus the decrease at A_(340nm)) was continuously monitoredspectrophometrically. The reaction mixture (100 μl) contained Abl kinase(4.4 nM. M315I Abl from deCode Genetics), peptide substrate(EAIYAAPFAKKK, 0.2 mM), MgCl₂ (10 mM), pyruvate kinase (4 units),lactate dehydrogenase (0.7 units), phosphoenol pyruvate (1 mM), and NADH(0.28 mM) in 90 mM Tris buffer containing 0.2% octyl-glucoside and 1%DMSO, pH 7.5. Test compounds were incubated with T315I Abl (SEQ ID NO:2)and other reaction reagents at 30° C. for 1 h before ATP (500 μM) wasadded to start the reaction. The absorption at 340 nm was monitoredcontinuously for 2 hours at 30° C. on Polarstar Optima plate reader(BMG). The reaction rate was calculated using the 1.0 to 2.0 h timeframe. Percent inhibition was obtained by comparison of reaction ratewith that of a control (i.e. with no test compound). IC₅₀ values werecalculated from a series of percent inhibition values determined at arange of inhibitor concentrations using software routines as implementedin the GraphPad Prism software package.

Abl T3151 kinase (SEQ ID NO: 2)GTSMDPSSPNYDKWEMERTDITMKHKLGGGQYGEVYEGVWKKYSLTVAVKTLKEDTMEVEEFLKEAAVMKEIKHPNLVQLLGVCTREPPFYIIIEFMTYGNLLDYLRECNRQEVNAVVLLYMATQISSAMEYLEKKNFIHRDLAARNCLVGENHLVKVADFGLSRLMTGDTYTAHAGAKFPIKWTAPESLAYNKFSIKSDVWAFGVLLWEIATYGMSPYFGIDLSQVYELLEKDYRMERPEGCPEKVYELMRACWQWNPSDRPSFAEIHQAFETMFQE BCR-Abl p210-e14a2 (SEQ ID NO: 3)MVDPVGFAEAWKAQFPDSEPPRMELRSVGDIEQELERCKASIRRLEQEVNQERFRMIYLQTLLAKEKKSYDRQRWGFRRAAQAFDGASEPRASASRPQPAPADGADPPPAEEPEARPDGEGSPGKARPGTARRPGAAASGERDDRGPPASVAALRSNFERIRKGHGQPGADAEKPFYVNVEFHHERGLVKVNDKEVSDRISSLGSQAMQMERKKSQHGAGSSVGDASRPPYRGRSSESSCGVDGDYEDAELNPRFLKDNLIDANGGSRPPWPPLEYQPYQSIYVGGIMEGEGKGPLLRSQSTSEQEKRLTWPRRSYSPRSFEDCGGGYTPDCSSNENLTSSEEDFSSGQSSRVSPSPTTYRMFRDKSRSPSQNSQQSFDSSSPPTPQCHKRHRHCPVVVSEATIVGVRKTGQIWPNDDEGAFHGDADGSFGTPPGYGCAADRAEEQRRHQDGLPYIDDSPSSSPHLSSKGRGSRDALVSGALKSTKASELDLEKGLEMRKWVLSGILASEETYLSHLEALLLPMKPLKAAATTSQPVLTSQQIETIFFKVPELYEIHKESYDGLFPRVQQWSHQQRVGDLFQKLASQLGVYRAFVDNYGVAMEMAEKCCQANAQFAEISENLRARSNKDAKDPTTKNSLETLLYKPVDRVTRSTLVLHDLLKHTPASHPDHPLLQDALRISQNFLSSINEEITPRRQSMTVKKGEHRQLLKDSFMVELVEGARKLRHVFLFTDLLLCTKLKKQSGGKTQQYDCKWYIPLTDLSFQMVDELEAVPNIPLVPDEELDALKIKISQIKSDIQREKRANKGSKATERLKKKLSEQESLLLLMSPSMAFRVHSRNGKSYTFLISSDYERAEWRENIREQQKKCFRSFSLTSVELQMLTNSCVKLQTVHSIPLTINKEDDESPGLYGFLNVIVHSATGFKQSSKALQRPVASDFEPQGLSEAARWNSKENLLAGPSENDPNLFVALYDFVASGDNTLSITKGEKLRVLGYNHNGEWCEAQTKNGQGWVPSNYITPVNSLEKHSWYHGPVSRNAAEYPLSSGINGSFLVRESESSPSQRSISLRYEGRVYHYRINTASDGKLYVSSESRFNTLAELVHHHSTVADGLITTLHYPAPKRNKPTVYGVSPNYDKWEMERTDITMKHKLGGGQYGEVYEGVWKKYSLTVAVKTLKEDTMEVEEFLKEAAVMKEIKHPNLVQLLGVCTREPPFYIITEFMTYGNLLDYLRECNRQEVNAVVLLYMATQISSAMEYLEKKNFIHRDLAARNCLVGENHLVKVADFGLSRLMTGDTYTAHAGAKFPIKWTAPESLAYNKFSIKSDVWAFGVLLWEIATYGMSPYPGIDRSQVYELLEKDYRMKRPEGCPEKVYELMRACWQWNPSDRPSFAEIHQAFETMFQESSISDEVEKELGKQGVRGAVTTLLQAPELPTKTRTSRRAAEHRDTTDVPEMPHSKGQGESDPLDHEPAVSPLLPRKERGPPEGGLNEDERLLPKDKKTNLFSALIKKKKKTAPTPPKRSSSFREMDGQPERRGAGEEEGRDISNGALAFTPLDTADPAKSPKPSNGAGVPNGALRESGGSGFRSPHLWKKSSTLTSSRLATGEEEGGGSSSKRFLRSCSVSCVPHGAKDTEWRSVTLPRDLQSTGRQFDSSTFGGHKSEKPALPRKRAGENRSDQVTRGTVTPPPRLVKKNEEAADEVFKDIMESSPGSSPPNLTPKPLRRQVTVAPASGLPHKEEAWKGSALGTPAAAEPVTPTSKAGSGAPRGTSKGPAEESRVRRHKHSSESPGRDKGKLSKLKPAPPPPPAASAGKAGGKPSQRPGQEAAGEAVLGAKTKATSLVDAVNSDAAKPSQPAEGLKKPVLPATFKPHPAKPSGTPISPAPVPLSTLPSASSALAGDQPSSTAFIPLISTRVSLRKTRQPPERASGAITKGVVLDSTEALCLAISGNSEQMASHSAVLEAGKNLYTFCVSYVDSIQQMRNKFAFREAINKLENNLRELQICPASAGSGPAATQDFSKLLSSVKEISDIVQR BCR-Abl p210-e13a2 (SEQ ID NO: 4)MVDPVGFAEAWKAQFPDSEPPRMELRSVGDIEQELERCKASIRRLEQEVNQERFRMIYLQTLLAKEKKSYDRQRWGFRRAAQAPDGASEPRASASRPQPAPADGADPPPAEEPEARPDGEGSPGKARPGTARRPGAAASGERDDRGPPASVAALRSNFERIRKGHGQPGADAEKPFYVNVEFHHERGLVKVNDKEVSDRISSLGSQAMQMERKKSQHGAGSSVGDASRPPYRGRSSESSCGVDGDYEDAELNPRFLKDNLIDANGGSRPPWPPLEYQPYQSIYVGGIMEGEGKGPLLRSQSTSEQEKRLTWPRRSYSPRSFEDCGGGYTPDCSSNENLTSSEEDFSSGQSSRVSPSPTTYRMFRDKSRSPSQNSQQSFDSSSPPTPQCHKRHRHCPVVVSEATIVGVRKTGQIWPNDDEGAFHGDADGSFGTPPGYGCAADRAEEQRPHQDGLPYIDDSPSSSPHLSSKGRGSRDALVSGALKSTKASELDLEKGLEMRKWVLSGILASEETYLSHLEALLLPMKPLKAAATTSQPVLTSQQIETIFFKVPELYEIHKESYDGLFPRVQQWSHQQRVGDLFQKLASQLGVYRAFVDNYGVAMEMAEKCCQANAQFAEISENLRARSNKDAKDPTTKNSLETLLYKPVDRVTRSTLVLHDLLKHTPASHPDHPLLQDALRISQNFLSSINEEITPRRQSMTVKKGEHRQLLKDSFMVELVEGARKLRHVFLFTDLLLCTKLKKQSGGKTQQYDCKWYIPLTDLSFQMVDELEAVPNIPLVPDEELDALKIKISQIKSDIQREKRANKGSKATERLKKKLSEQESLLLLMSPSMAFRVHSRNGKSYTFLISSDYERAEWRENIREQQKKCFRSFSLTSVELQMLTNSCVKLQTVHSIPETINKEEALQRPVASDFEPQGLSEAARWNSKENLLAGFSENDPNLFVALYDFVASGDNTLSITKGEKLRVLGYNHNGEWCEAQTKNGQGWVPSNYITPVNSLEKHSWYHGPVSRNAAEYPLSSGINGSFLVRESESSPSQRSISLRYEGRVYHYRINTASDGKLYVSSESRFNTLAELVHHHSTVADGLITTLHYPAPKRNKPTVYGVSPNYDKWEMERTDITMKHKLGGGQYGEVYEGVWKKYSLTVAVKTLKEDTMEVEEFLKEAAVMKEIKHPNLVQLLGVCTREPPFYIITEFMTYGNLLDYLRECNRQEVNAVVLLYMATQISSAMEYLEKKNFIHRDLAARNCLVGENHLVKVADFGLSRLMTGDTYTAHAGAKFPIKWTAPESLAYNKFSIKSDVWAFGVLLWEIATYGMSPYPGIDRSQVYELLEKDYRMKRPEGCPEKVYELMRACWQWNPSDRPSFAEIHQAFETMFQESSISDEVEKELGKQGVRGAVTTLLQAPELPTKTRTSRRAAEHRDTTDVPEMPHSKGQGESDPLDHEPAVSPLLPRKERGPPEGGLNEDERLLPKDKKTNLFSALIKKKKKTAPTPPKRSSSFREMDGQPERRGAGEEEGRDISNGALAFTPLDTADPAKSPKPSNGAGVPNGALRESGGSGFRSPHLWKKSSTLTSSRLATGEEEGGGSSSKRFLRSCSVSCVPHGAKDTEWRSVTLPRDLQSTGRQFDSSTFGGHKSEKPALPRKRAGENRSDQVTRGTVTPPPRLVKKNEEAADEVFKDIMESSPGSSPPNLTPKPLRRQVTVAPASGLPHKEEAWKGSALGTPAAAEPVTPTSKAGSGAPRGTSKGPAEESRVRRHKHSSESPGRDKGKLSKLKPAPPPPPAASAGKAGGKPSQRPGQEAAGEAVLGAKTKATSLVDAVNSDAAKPSQPAEGLKKPVLPATPKPHPAKPSGTPISPAPVPLSTLPSASSALAGDQPSSTAFIPLISTRVSLRKTRQPPERASGAITKGVVLDSTEALCLAISGNSEQMASHSAVLEAGKNLYTFCVSYVDSIQQMRNKFAFREAINKLENNLRELQICPASAGSGPAATQDFSKLLSSVKEI SDIVQR BCR-Ablp190-e1a2 (SEQ ID NO: 5)MVDPVGFAEAWKAQFPDSEPPRMELRSVGDIEQELERCKASIRRLEQEVNQERFRMIYLQTLLAKEKKSYDRQRWGFRRAAQAPDGASEPRASASRPQPAPADGADPPPAEEPEARPDGEGSPGKARPGTARRPGAAASGERDDRGPPASVAALRSNFERIRKGHGQPGADAEKPFYVNVEFHHERGLVKVNDKEVSDRISSLGSQAMQMERKKSQHGAGSSVGDASRPPYRGRSSESSCGVDGDYEDAELNPRFLKDNLIDANGGSRPPWPPLEYQPYQSIYVGGIMEGEGKGPLLRSQSTSEQEKRLTWPRRSYSPRSFEDCGGGYTPDCSSNENLTSSEEDFSSGQSSRVSPSPTTYRMFRDKSRSPSQNSQQSFDSSSPPTPQCHKRHRHCPVVVSEATIVGVRKTGQIWPNDDEGAFHGDAEALQRPVASDFEPQGLSEAARWNSKENLLAGPSENDPNLFVALYDFVASGDNTLSITKGEKLRVLGYNHNGEWCEAQTKNGQGWVPSNYITPVNSLEKHSWYHGPVSRNAAEYPLSSGINGSFLVRESESSPSQRSISLRYEGRVYHYRINTASDGKLYVSSESRFNTLAELVHHHSTVADGLITTLHYPAPKRNKPTVYGVSPNYDKWEMERTDITMKHKLGGGQYGEVYEGVWKKYSLTVAVKTLKEDTMEVEEFLKEAAVMKEIKHPNLVQLLGVCTREPPFYIITEFMTYGNLLDYLRECNRQEVNAVVLLYMATQISSAMEYLEKKNFIHRDLAARNCLVGENHLVKVADFGLSRLMTGDTYTAHAGAKFPIKWTAPESLAYNKFSIKSDVWAFGVLLWEIATYGMSPYPGIDRSQVYELLEKDYRMKRPEGCPEKVYELMRACWQWNPSDRPSFAEIHQAFETMFQESSISDEVEKELGKQGVRCAVTTLLQAPELPTKTRTSRRAAEHRDTTDVPEMPHSKGQGESDPLDHEPAVSPLLPRKERGPPEGGLNEDERLLPKDKKTNLFSALIKKKKKTAPTPPKRSSSFREMDGQPERRGAGEEEGRDISNGALAFTPLDTADPAKSPKPSNGAGVPNGALRESGGSGFRSPHLWKKSSTLTSSRLATGEEEGGGSSSKRFLRSCSVSCVPHGAKDTEWRSVTLPRDLQSTGRQFDSSTFGGHKSEKPALFRKRAGENRSDQVTRGTVTPPPRLVKKNEEAADEVFKDIMESSPGSSPPNLTPKPLRRQVTVAPASGLPHKREAWKGSALGTPAAAEPVTPTSKAGSGAPRGTSKGPAEESRVRRHKHSSESPGRDRGKLSKLKPAPPPPPAASAGKAGGKPSQRPGQEAAGEAVLGAKTKATSLVDAVNSDAAKPSQPAEGLKKPVLPATPKPHPAKPSGTPISPAPVPLSTLPSASSALAGDQPSSTAFIPLISTRVSLRKTRQPPERASGAITKGVVLDSTEALCLAISGNSEQMASHSAVLEAGKNLYTFCVSYVDSIQQMRNKFAFREAINKLENNLRELQICPASAGSGPAATQDFSKLLSSVKEISDIVQR BCR-Abl p210-e14a2 T315I (SEQ ID NO: 6)MVDPVGFAEAWKAQFPDSEPPRMELRSVGDIEQELERCKASIRRLEQEVNQERFRMIYLQTLLAKEKKSYDRQRWGFRRAAQAPDGASEPRASASRPQPAPADGADPPPAEEPEARPDGEGSPGKARPGTARRPGAAASGERDDRGPPASVAALRSNFERIRKGHGQPGADAEKPFYVNVEFHHERGLVKVNDKEVSDRISSLGSQAMQMERKKSQHGAGSSVGDASRPPYRGRSSESSCGVDGDYEDAELNPRFLKDNLIDANGGSRPPWPPLEYQPYQSIYVGGIMEGEGKGPLLRSQSTSEQEKRLTWPRRSYSPRSFEDCGGGYTPDCSSNENLTSSEEDFSSGQSSRVSPSPTTYRMFRDKSRSPSQNSQQSFDSSSPPTPQCHKRHRHCPVVVSEATIVGVRKTGQIWPNDDEGAFHGDADGSFGTPPGYGCAADRAEEQRRHQDGLPYIDDSPSSSPHLSSKGRGSRDALVSGALKSTKASELDLEKGLEMRKWVLSGILASEETYLSHLEALLLPMKPLKAAATTSQPVLTSQQIETIFFKVPELYEIHKESYDGLFPRVQQWSHQQRVGDLPQKLASQLGVYRAFVDNYGVAMEMAEKCCQANAQFAEISENLRARSNKDAKDPTTKNSLETLLYKPVDRVTRSTLVLHDLLKHTPASHPDHPLLQDALRISQNFLSSINEEITPRRQSMTVKKGEHRQLLKDSFMVELVEGARKLRHVFLFTDLLLCTKLKKQSGGKTQQYDCKWYIPLTDLSFQMVDELEAVPNIPLVPDEELDALKIKISQIKSDIQREKRANKGSKATERLKKKLSEQESLLLLMSPSMAFRVHSRNGKSYTFLISSDYERAEWRENIREQQKKCFRSFSLTSVELQMLTNSCVKLQTVHSIPLTINKEDDESPGLYGFLNVIVHSATGFKQSSKALQRPVASDFEPQGLSEAARWNSKENLLAGPSENDPNLFVALYDFVASGDNTLSITKGEKLRVLGYNHNGEWCEAQTKNGQGWVPSNYITPVNSLEKHSWYHGPVSRNAAEYPLSSGINGSFLVRESESSPSQRSISLRYEGRVYHYRINTASDGKLYVSSESRFNTLAELVHHHSTVADGLITTLHYPAPKRNKPTVYGVSPNYDKWEMERTDITMKHKLGGGQYGEVYEGVWKKYSLTVAVKTLKEDTMEVEEFLKEAAVMKEIKHPNLVQLLGVCTREPPFYIIIEFMTYGNLLDYLRECNRQEVNAVVLLYMATQISSAMEYLEKKNFIHRDLAARNCLVGENHLVKVADFGLSRLMTGDTYTAHAGAKFPIKWTAPESLAYNKFSIKSDVWAFGVLLWEIATYGMSPYPGIDRSQVYELLEKDYRMKRPEGCPEKVYELMRACWQWNPSDRPSFAEIHQAFETMFQESSISDEVEKELGKQGVRGAVTTLLQAPELPTKTRTSRRAAEHRDTTDVPEMPHSKGQGESDPLDHEPAVSPLLPRKERGPPEGGLNEDERLLPKDKKTNLFSALIKKKKKTAPTPPKRSSSFREMDGQPERRGAGEEEGRDISNGALAFTPLDTADPAKSPKPSNGAGVPNGALRESGGSGFRSPHLWKKSSTLTSSRLATGEEEGGGSSSKRFLRSCSVSCVPHGAKDTEWRSVTLPRDLQSTGRQFDSSTFGGHKSEKPALPRKRAGENRSDQVTRGTVTPPPRLVKKNEEAADEVFKDIMESSPGSSPPNLTPKPLRRQVTVAPASGLPHKEEAWKGSALGTPAAAEPVTPTSKAGSGAPRGTSKGPAEESRVRRHKHSSESPGRDKGKLSKLKPAPPPPPAASAGKAGGKPSQRPGQEAAGEAVLGAKTKATSLVDAVNSDAAKPSQPAEGLKKPVLPATPKPHPAKPSGTPISPAPVPLSTLPSASSALAGDQPSSTAFIPLISTRVSLRKTRQPPERASGAITKGVVLDSTEALCLAISGNSEQMASHSAVLEAGKNLYTFCVSYVDSIQQMRNKFAFREAINKLENNLRELQICPASAGSGFAATQDFSKLLSSVKEISDIVQR BCR-Abl p210-e13a2 T315I (SEQ ID NO: 7)MVDPVGFAEAWKAQFPDSEPPRMELRSVGDIEQELERCKASIRRLEQEVNQERFRMIYLQTLLAKEKKSYDRQRWGFRRAAQAPDGASEPRASASRPQPAPADGADPPPAEEPEARPDGEGSPGKARPGTARRPGAAASGERDDRGPPASVAALRSNFERIRKGHGQPGADAEKPFYVNVEFHHERGLVKVNDKEVSDRISSLGSQAMQMERKKSQHGAGSSVGDASRPPYRGRSSESSCGVDGDYEDAELNPRFLKDNLIDANGGSRPPWPPLEYQPYQSIYVGGIMEGEGKGPLLRSQSTSEQEKRLTWPRRSYSPRSFEDCGGGYTPDCSSNENLTSSEEDFSSGQSSRVSPSPTTYRMFRDKSRSPSQNSQQSFDSSSPPTPQCHKPHRHCPVVVSEATIVGVRKTGQIWPNDDEGAFHGDADGSFGTPPGYGCAADRAEEQRRHQDGLPYIDDSPSSSPHLSSKGRGSRDALVSGALKSTKASELDLEKGLEMRKWVLSGILASEETYLSHLEALLLPMKPLKAAATTSQPVLTSQQIETIFFKVPELYEIHKESYDGLFPRVQQWSHQQRVGDLFQKLASQLGVYRAFVDNYGVAMEMAEKCCQANAQFAEISENLRARSNKDAKDPTTKNSLETLLYKPVDRVTRSTLVLHDLLKHTPASHPDHPLLQDALRISQNFLSSINEEITPRRQSMTVKKGEHRQLLKDSFMVELVEGARKLRHVFLFTDLLLCTKLKKQSGGKTQQYDCKWYIPLTDESFQMVDELEAVPNIPLVPDEELDALKIKISQIKSDIQREKRANKGSKATERLKKKLSEQESLLLLMSPSMAFRVHSRNGKSYTFLISSDYERAEWRENIREQQKKCFRSPSLTSVELQMLTNSCVKLQTVHSIPLTINKEEALQRPVASDFEPQGLSEAAPWNSKENLLAGPSENDPNLFVALYDFVASQDNTLSITKGEKLRVLGYNHNGEWCEAQTKNGQGWVPSNYITPVNSLEKHSWYHGPVSRNAAEYPLSSGINGSFLVRESESSPSQRSISLRYEGRVYHYRINTASDGKLYVSSESRFNTLAELVHHHSTVADGLITTLHYPAPKRNKPTVYGVSPNYDKWEMERTDITMKHKLGGGQYGEVYEGVWKKYSLTVAVKTLKEDTMEVEEFLKEAAVMKEIKHPNLVQLLGVCTREPPFYIIIEFMTYGNLLDYLRECNRQEVNAVVLLYMATQISSAMEYLEKKNFIHRDLAARNCLVGENHLVKVADFGLSRLMTGDTYTAHAGAKFPIKWTAFESLAYNKFSIKSDVWAFGVLLWEIATYGMSPYPGIDRSQVYELLEKDYRMKRPEGCPEKVYELMRACWQWNPSDRPSFAEIHQAFETMFQESSISDEVEKELGKQGVRGAVTTLLQAPELPTKTRTSRRAAEHRDTTDVPEMPHSKGQGESDPLDHEPAVSPLLPRKERGPFEGGLNEDERLLPKDKKTNLFSALIKKKKKTAPTPPKRSSSFREMDGQFERRGAGEEEGRDISNGALAFTPLDTADPAKSFKPSNGAGVPNGALRESGGSGFRSPHLWKKSSTLTSSRLATGEEEGGGSSSKRFLRSCSVSCVPHGAKDTEWRSVTLPRDLQSTGRQFDSSTFGGHKSEKPALPRKRAGENRSDQVTRGTVTPPPRLVKKNEEAADEVFKDIMESSPGSSPPNLTPKPLRRQVTVAPASGLPHKEEAWKGSALGTPAAAEPVTPTSKAGSGAPRGTSKGPAEESRVRRHKHSSESPGRDKGKLSKLKPAPPPPPAASAGKAGGKPSQRPGQEAAGEAVLGAKTKATSLVDAVNSDAAKPSQPAEGLKKPVLPATPKPHPAKPSGTPISPAPVPLSTLPSASSALAGDQPSSTAFIPLISTRVSLRKTRQPPERASGAITKGVVLDSTEALCLAISGNSEQMASHSAVLEAGKNLYTFCVSYVDSIQQMRNKFAFREAINKLENNLRELQICPASAGSGPAATQDFSKLLSSVKEI SDIVQR BCR-Ablp190-e1a2 (SEQ ID NO: 8)MVDPVGFAEAWKAQFPDSEPPRMELRSVGDIEQELERCKASIRRLEQEVNQERFRMIYLQTLLAKEKKSYDRQRWGFRRAAQAFDGASEPRASASRPQPAPADGADPPPAEEPEARPDGEGSPGKARPGTARRPQAAASGERDDRGPPASVAALRSNFERIRKGHGQPGADAEKPFYVNVEFHHERGLVKVNDKEVSDRISSLGSQAMQMERKKSQHGAGSSVGDASRPPYRGRSSESSCGVDGDYEDAELNPRFLKDNLIDANGGSRPPWFPLEYQPYQSIYVGGIMEGEGKGPLLRSQSTSEQEKRLTWPRRSYSPRSFEDCGGGYTPDCSSNENLTSSEEDFSSGQSSRVSPSPTTYRMFRDKSRSPSQNSQQSFDSSSPPTPQCHKRHRHCPVVVSEATIVGVRKTGQIWPNDDEGAFHGDAEALQRPVASDFEPQGLSEAARWNSKENLLAGPSENDPNLFVALYDFVASGDNTLSITKGEKLRVLGYNHNGEWCEAQTKNGQGWVPSNYITPVNSLEKHSWYHGPVSRNAAEYPLSSGINGSFLVRESESSPSQRSISLRYEGRVYHYRINTASDGKLYVSSESRFNTLAELVHHHSTVADGLITTLHYPAPKRNRPTVYGVSPNYDKWEMERTDITMKHKLGGGQYGEVYEGVWKKYSLTVAVKTLKEDTMEVEEFLKEAAVMKEIKHPNLVQLLGVCTREPPFYIIIEFMTYGNLLDYLRECNRQEVNAVVLLYMATQISSAMEYLEKKNFIHRDLAARNCLVGENHLVKVADFGLSRLMTGDTYTAHAGAKFPIKWTAPESLAYNKFSIKSDVWAFGVLLWEIATYGMSPYPGIDRSQVYELLEKDYRMKRPEGCPEKVYELMPACWQWNPSDRPSFAEIHQAFETMFQESSISDEVEKELGKQGVRGAVTTLLQAPELPTKTRTSRRAAEHRDTTDVPEMPHSKGQGESDPLDHEPAVSFLLPRKERGPPEGGLNEDERLLPKDKKTNLFSALIKKKKKTAPTPPKRSSSFREMDGQPERRGAGEEEGRDISNGALAFTPLDTADPAKSPKPSNGAGVPNGALRESGGSGFRSPHLWKKSSTLTSSRLATGEEEGGGSSSKRFLRSCSVSCVPHGAKDTEWRSVTLPRDLQSTGRQFDSSTFGGHKSEKPALPRKRAGENRSDQVTRGTVTPPPRLVKKNEEAADEVFKDIMESSPGSSPPNLTPKPLRRQVTVAPASGLPHKEEAWKGSALGTPAAAEPVTPTSKAGSGAPRGTSKGPAEESRVRRHKHSSESPGRDKGKLSKLKPAPPPPPAASAGKAGGKPSQRPGQEAAGEAVLGAKTKATSLVDAVNSDAAKPSQPAEGLKKPVLPATPKPHPAKPSGTPISPAPVPLSTLPSASSALAGDQPSSTAFIPLISTRVSLRKTRQPPERASGAITKGVVLDSTEALCLAISGNSEQMASHSAVLEAGKNLYTFCVSYVDSIQQMRNKFAFREAINKLENNLRELQICPASAGSGPAATQDFSKLLSSVKEISDIVQR C-Kit kinase (SEQ ID NO: 9) assay

Activity of c-Kit kinase (SEQ ID NO:9) was determined by following theproduction of ADP from the kinase reaction through coupling with thepyruvate kinase/lactate dehydrogenase system (e.g.. Schindler, et al.Science (2000) 289, 1938-1942). In this assay, the oxidation of NADH(thus the decrease at A340 nm) was continuously monitoredspectrophometrically. The reaction mixture (100 μl) contained c-Kit(cKIT residues T544-V976, from ProQinase, 5.4 nM), polyE4Y (1 mg/ml),MgCl2 (10 mM), pyruvate kinase (4 units), lactate dehydrogenase (0.7units), phosphoenol pyruvate (1 mM), and NADH (0.28 mM) in 90 mM Trisbuffer containing 0.2% octyl-glucoside and 1% DMSO, pH 7.5. Testcompounds were incubated with C-Met (SEQ ID NO:9) and other reactionreagents at 22° C. for <2 min before ATP (200 μM) was added to start thereaction. The absorption at 340 nm was monitored continuously for 0.5hours at 30° C. on Polarstar Optima plate reader (BMG). The reactionrate was calculated using the 0 to 0.5 h time frame. Percent inhibitionwas obtained by comparison of reaction rate with that of a control (i.e.with no test compound). IC50 values were calculated from a series ofpercent inhibition values determined at a range of inhibitorconcentrations using software routines as implemented in the GraphPadPrism software package.

c-Kit with N-terminal GST fusion (SEQ ID NO: 9)LGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVDIRYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIWPLQGWQATFGGGDHPPKSDLVPRHNQTSLYKKAGSAAAVLEENLYFQGTYKYLQKPMYEVQWKVVEEINGNNYVYIDPTQLPYDHKWEFPRNRLSFGKTLGAGAFGKVVEATAYGLIKSDAAMTVAVKMLKPSAHLTEREALMSELKVLSYLGNHMNIVNLLGACTTGGPTLVITEYCCYGDLLNFLRRKRDSFICSKQEDHAEAALYKNLLHSKESSCSDSTNEYMDMKPGVSYVVPTKADKRRSVRIGSYIERDVTPAIMEDDELALDLEDLLSFSYQVAKGMAFLASKNCIHRDLAARNILLTHGRITKICDFGLARDIKNDSNYVVKGNARLPVKWMAPESIFNCVYTFESDVWSYGIFLWELFSLGSSPYPGMPVDSKFYKMIKEGFRMLSFEHAPAEMYDIMKTCWDADPLKRPTFKQIVQLIEKQISESTNHIYSNLANCSPNRQKPVVDHSVRINSVGSTASSSQPLLVHDDV

C-Met Kinase (SEQ ID NO:10) Assay

Activity of C-Met kinase (SEQ ID NO:10) was determined by following theproduction of ADP from the kinase reaction through coupling with thepyruvate kinase/lactate dehydrogenase system (e.g., Schindler, et al.Science (2000) 289, 1938-1942). In this assay, the oxidation of NADH(thus the decrease at A340 nm) was continuously monitoredspectrophometrically. The reaction mixture (100 μl) contained C-Met(c-Met residues: 956-1390, from Invitrogen, catalogue #PV3143, 6 nM),polyE4Y (1 mg/ml), MgCl2 (10 mM), pyruvate kinase (4 units), lactatedehydrogenase (0.7 units), phosphoenol pyruvate (1 mM), and NADH (0.28mM) in 90 mM Tris buffer containing 0.25 mM DTT, 0.2% octyl-glucosideand 1% DMSO, pH 7.5. Test compounds were incubated with C-Met (SEQ IDNO:10) and other reaction reagents at 22° C. for 0.5 h before ATP (100μM) was added to start the reaction. The absorption at 340 nm wasmonitored continuously for 2 hours at 30° C. on Polarstar Optima platereader (BMG). The reaction rate was calculated using the 1.0 to 2.0 htime frame. Percent inhibition was obtained by comparison of reactionrate with that of a control (i.e. with no test compound), IC50 valueswere calculated from a series of percent inhibition values determined ata range of inhibitor concentrations using software routines asimplemented in the GraphPad Prism software package.

cMet Kinase (SEQ ID NO: 10)MSYYHHHHHHDYDIPTTENLYFQGAMLVPRGSPWIPFTMKKRKQIKDLGSELVRYDARVHTPHLDRLVSARSVSPTTEMVSNESVDYRATFPEDQFPNSSQNGSCRQVQYPLTDMSPILTSGDSDISSPLLQNTVHIDLSALNPELVQAVQHVVIGPSSLIVHFNEVIGRGHFGCVYHGTLLDNDGKKIHCAVKSLNRITDIGEVSQFLTEGIIMKDFSHPNVLSLLGICLRSECSPLVVLPYMKHGDLRNFIRNETHNPTVKDLIGFGLQVAKGMKYLASKKFVHRDLAARNCMLDEKFTVKVADFGLARDMYDKEYYSVHNKTGAKLPVKWMALESLQTQKFTTKSDVWSFGVLLWELMTRGAPPYPDVNTFDITVYLLQGRRLLQPEYCPDPLYEVMLKCWHPKAEMRPSFSELVSRISAIFSTFIGEHYVHVNATYVNVKCVAPYPSLLSSEDNADDEVDTRPASFWETS

The biochemical IC₅₀ Values of other compounds disclosed herein are atleast 10 μM against Abl enzyme.

Cell Culture

BaF3 cells (parental or transfected with the following: wild type p210BCR-Abl and T315I p210 BCR-Abl was obtained from Professor Richard VanEtten (New England Medical Center, Boston, Mass.). Briefly, cells weregrown in RPMI 1640 supplemented with 10% characterized fetal bovineserum (HyClone, Logan, Utah) at 37 degrees Celsius, 5% CO₂, 95%humidity. Cells were allowed to expand until reaching 80% saturation atwhich point they were subcultured or harvested for assay use.

Cell Proliferation Assay

A serial dilution of test compound was dispensed into a 96 well blackclear bottom plate (Coming, Coming, N.Y.). For each cell line, threethousand cells were added per well in complete growth medium. Plateswere incubated for 72 hours at 37 degrees Celsius, 5% CO₂, 95% humidity.At the end of the incubation period Cell Titer Blue (Promega, Madison,Wis.) was added to each well and an additional 4.5 hour incubation at 37degrees Celsius, 5% CO₂, 95% humidity was performed. Plates were thenread on a BMG Fluostar Optima (BMG, Durham, N.C.) using an excitation of544 nM and an emission of 612 nM. Data was analyzed using Prism software(Graphpad, San Diego. Calif.) to calculate IC50's.

1. Compounds of the formula Ia

and wherein the pyridine ring may be optionally substituted with one ormore R20 moieties; each D is individually taken from the groupconsisting of C, CH, C—R20, N-Z3, N, O and S, such that the resultantring is taken from the group consisting of triazolyl, isoxazolyl,isothiazolyl, oxazolyl, and thiadiazolyl; wherein E is selected from thegroup consisting of phenyl, pyridyl, and pyrimidinyl; E may beoptionally substituted with one or two R16 moieties; wherein A is a ringsystem selected from the group consisting of phenyl, naphthyl,cyclopentyl, cyclohexyl, G1, G2, and G3; G1 is a heteroaryl taken fromthe group consisting of pyrrolyl, furyl, thienyl, oxazolyl, thiazolyl,isoxazol-4-yl, isoxazol-5-yl, isothiazolyl, imidazolyl, pyrazolyl,oxadiazolyl, thiadiazolyl, triazolyl, tetrazolyl, pyrazinyl,pyridazinyl, triazinyl, pyridinyl, and pyrimidinyl; G2 is a fusedbicyclic heteroaryl taken from the group consisting of indolyl,indolinyl, isoindolyl, isoindolinyl, indazolyl, benzofuranyl,benzothienyl, benzothiazolyl, benzothiazolonyl, benzoxazolyl,benzoxazolonyl, benzisoxazolyl, benzisothiazolyl, benzimidazolyl,benzimidazolonyl, benztriazolyl, imidazopyridinyl, pyrazolopyridinyl,imidazolonopyridinyl, thiazolopyridinyl, thiazolonopyridinyl,oxazolopyridinyl, oxazolonopyridinyl, isoxazolopyridinyl,isothiazolopyridinyl, triazolopyridinyl, imidazopyrimidinyl,pyrazolopyrimidinyl, imidazolonopyrmidinyl, thiazolopyridiminyl,thiazolonopyrimidinyl, oxazolopyridiminyl, oxazolonopyrimidinyl,isoxazolopyrimidinyl, isothiazolopyrimidinyl, triazolopyrimidinyl,dihydropurinonyl, pyrrolopyrimidinyl, purinyl, pyrazolopyrimidinyl,phthalimidyl, phthalimidinyl, pyrazinylpyridinyl, pyridinopyrimidinyl,pyrimidinopyrimidinyl, cinnolinyl, quinoxalinyl, quinazolinyl,quinolinyl, isoquinolinyl, phthalazinyl, benzodioxyl,benzisothiazoline-1,1,3-trionyl, dihydroquinolinyl,tetrahydroquinolinyl, dihydroisoquinolyl, tetrahydroisoquinolinyl,benzoazepinyl, benzodiazepinyl, benzoxapinyl, and benzoxazepinyl; G3 isa heterocyclyl taken from the group consisting of oxetanyl, azetadinyl,tetrahydrofuranyl, pyrrolidinyl, oxazolinyl, oxazolidinyl, imidazolonyl,pyranyl, thiopyranyl, tetrahydropyranyl, dioxalinyl, piperidinyl,morpholinyl, thiomorpholinyl, thiomorpholinyl S-oxide, thiomorpholinylS-dioxide, piperazinyl, azepinyl, oxepinyl, diazepinyl, tropanyl, andhomotropanyl; the A ring may be optionally substituted with one or twoR2 moieties; X is selected from the group consisting of —O—,—S(CH₂)_(n)—, —N(R3)(CH₂)_(n)—, —(CH₂)_(p)—, and wherein the carbonatoms of —(CH₂)_(n)—, —(CH₂)_(p)—, of X may be further substituted byoxo or one or more C1-C6alkyl moieties; when A, G1, G2 or G3 has one ormore substitutable sp2-hybridized carbon atoms, each respective sp2hybridized carbon atom may be optionally substituted with a Z1substituent; when A, G1, G2 or G3 has one or more substitutablesp3-hybridized carbon atoms, each respective sp3 hybridized carbon atommay be optionally substituted with a Z2 substituent; when A, G1, G2 orG3 has one or more substitutable nitrogen atoms, each respectivenitrogen atom may be optionally substituted with a Z4 substituent; eachZ1 is independently and individually selected from the group consistingof C1-6alkyl, branched C3-C7alkyl, C3-C8cycloalkyl, halogen,fluoroC1-C6alkyl wherein the alkyl moiety can be partially or fullyfluorinated, cyano, C1-C6alkoxy, fluoroC1-C6alkoxy wherein the alkylmoiety can be partially or fully fluorinated, —(CH₂)_(n)OH, oxo,C1-C6alkoxyC1-C6alkyl, (R4)₂N(CH₂)_(n)—, (R3)₂N(CH₂)_(n)—,(R4)₂N(CH₂)_(q)N(R4)(CH₂)_(n)—, (R4)₂N(CH₂)_(q)O(CH₂)_(n)—, (R3)₂NC(O)—,(R4)₂NC(O)—, (R4)₂NC(O)C1-C6alkyl-, —(R4)NC(O)R8, C1-C6alkoxycarbonyl-,-carboxyC1-C6alkyl, C1-C6alkoxycarbonylC1-C6alkyl-, (R3)₂NSO₂—, —SOR3,(R4)₂NSO₂—, —N(R4)SO₂R8, —O(CH₂)_(q)OC1-C6alkyl, —SO₂R3, —SOR4, —C(O)R8,—C(O)R6, —C(═NOH)R6, —C(═NOR3)R6, —(CH₂)_(n)N(R4)C(O)R8,—N(R3)(CH₂)_(q)O-alkyl, —N(R3)(CH₂)_(q)N(R4)₂, nitro, —CH(OH)CH(OH)R4,—C(═NH)N(R4)₂, —C(═NOR3)N(R4)₂, —NHC(═NH)R8, R17 substituted G3, R17substituted pyrazolyl and R17 substituted imidazolyl; in the event thatZ1 contains an alkyl or alkylene moiety, such moieties may be furthersubstituted with one or more C1-C6alkyls; each Z2 is independently andindividually selected from the group consisting of aryl, C1-C6alkyl,C3-C8cycloalkyl, branched C3-C7alkyl, hydroxyl, hydroxyC1-C6alkyl-,cyano, (R3)₂N—, (R4)₂N—, (R4)₂NC1-C6alkyl-,(R4)₂NC2-C6alkylN(R4)(CH₂)_(n)—, (R4)₂NC2-C6alkylO(CH₂)_(n)—,(R3)₂NC(O)—, (R4)₂NC(O)—, (R4)₂NC(O)—C1-C6alkyl-, carboxyl,-carboxyC1-C6alkyl, C1-C6alkoxycarbonyl-,C1-C6alkoxycarbonylC1-C6alkyl-, (R3)₂NSO₂—, (R4)₂NSO₂—, —SO₂R8,—(CH₂)_(n)N(R4)C(O)R8, —C(O)R8, ═O, ═NOH, and ═N(OR6); in the event thatZ2 contains an alkyl or alkylene moiety, such moieties may be furthersubstituted with one or more C1-C6alkyls; each Z3 is independently andindividually selected from the group consisting of H, C1-C6alkyl,branched C3-C7alkyl, C3-C8cycloalkyl, fluoroC1-C6alkyl wherein the alkylmoiety can be partially or fully fluorinated, hydroxyC2-C6alkyl-,C1-C6alkoxycarbonyl-, —C(O)R8, R5C(O)(CH₂)_(n)—, (R4)₂NC(O)—,(R4)₂NC(O)C1-C6alkyl-, R8C(O)N(R4)(CH₂)_(q)—, (R3)₂NSO₂—, (R4)₂NSO₂—,—(CH₂)_(q)N(R3)₂, and —(CH₂)_(q)N(R4)₂; each Z4 is independently andindividually selected from the group consisting of C1-C6alkyl, branchedC3-7alkyl, hydroxyC2-C6alkyl-, C1-C6alkoxyC2-C6alkyl-.(R4)₂N—C2-C6alkyl-, (R4)₂N—C2-C6alkylN(R4)-C2-C6alkyl-,(R4)₂N—C2-C6alkyl-O—C2-C6alkyl-(R4)₂NC(O)C1-C6alkyl-, carboxyC1-C6alkyl,C1-C6alkoxycarbonylC1-C6alkyl-, —C2-C6alkylN(R4)C(O)R8, R8-C(═NR3)—,—SO₂R8, and —COR8; in the event that Z4 contains an alkyl or alkylenemoiety, such moieties may be further substituted with one or moreC1-C6alkyls; each R2 is selected from the group consisting of H,C1-C6alkyl, branched C3-C8alkyl, R19 substituted C3-C8cycloalkyl-,fluoroC1-C6alkyl- wherein the alkyl is fully or partially fluorinated,halogen, cyano, C1-C6alkoxy-, and fluoroC1-C6alkoxy- wherein the alkylgroup is fully or partially fluorinated, hydroxyl substitutedC1-C6alkyl-, hydroxyl substituted branched C3-C8alkyl-, cyanosubstituted C1-C6alkyl-, cyano substituted branched C3-C8alkyl-,(R3)₂NC(O)C1-C6alkyl-, and (R3)₂NC(O)C3-C8 branched alkyl-; wherein eachR3 is independently and individually selected from the group consistingof H, C1-C6alkyl, branched C3-C7alkyl, and C3-C8cycloalkyl; each R4 isindependently and individually selected from the group consisting of H,C1-C6alkyl, hydroxyC1-C6alkyl-, dihydroxyC1-C6alkyl-,C1-C6alkoxyC1-C6alkyl-, branched C3-C7alkyl, branchedhydroxyC1-C6alkyl-, branched C1-C6alkoxyC1-C6alkyl-, brancheddihydroxyC1-C6alkyl-, —(CH₂)_(p)N(R7)₂, —(CH₂)_(p)C(O)N(R7)₂,—(CH₂)_(n)C(O)OR3, and R19 substituted C3-C8cycloalkyl-; each R5 isindependently and individually selected from the group consisting of

and wherein the symbol (##) is the point of attachment to Z3; each R6 isindependently and individually selected from the group consisting ofC1-C6alkyl, branched C3-C7alkyl, and R19 substituted C3-C8cycloalkyl-;each R7 is independently and individually selected from the groupconsisting of H, C1-C6alkyl, hydroxyC2-C6alkyl-, dihydroxyC2-C6alkyl-,C1-C6alkoxyC2-C6alkyl-, branched C3-C7alkyl, branchedhydroxyC2-C6alkyl-, branched C1-C6alkoxyC2-C6alkyl-, brancheddihydroxyC2-C6alkyl-, —(CH₂)_(n)C(O)OR3, R19 substitutedC3-C8cycloalkyl- and —(CH₂)_(n)R17; each R8 is independently andindividually selected from the group consisting of C1-C6alkyl, branchedC3-C7alkyl, fluoroC1-C6alkyl- wherein the alkyl moiety is partially orfully fluorinated, R19 substituted C3-C8cycloalkyl-, —OH, C1-C6alkoxy,—N(R3)₂, and —N(R4)₂; each R10 is independently and individuallyselected from the group consisting of —CO₂H, —CO₂C1-C6alkyl,—C(O)N(R4)₂, OH, C1-C6alkoxy and —N(R4)₂; each R16 is independently andindividually selected from the group consisting of H, C1-C6alkyl,branched C3-C7alkyl, R19 substituted C3-C8cycloalkyl-, halogen,fluoroC1-C6alkyl- wherein the alkyl moiety can be partially or fullyfluorinated, cyano, hydroxyl, C1-C6alkoxy, fluoroC1-C6alkoxy- whereinthe alkyl moiety can be partially or fully fluorinated, —N(R3)₂,—N(R4)₂, R3 substituted C2-C3alkynyl- and nitro; each R17 isindependently and individually selected from the group consisting of H,C1-C6alkyl, branched C3-C7alkyl, R19 substituted C3-C8cycloalkyl-,halogen, fluoroC1-C6alkyl- wherein the alkyl moiety can be partially orfully fluorinated, cyano, hydroxyl, C1-C6alkoxy, fluoroC1-C6alkoxy-wherein the alkyl moiety can be partially or fully fluorinated, —N(R3)₂,—N(R4)₂, and nitro; each R19 is independently and individually selectedfrom the group consisting of H, OH and C1-C6alkyl; each R20 isindependently and individually selected from the group consisting ofC1-C6alkyl, branched C3-C7alkyl, R19 substituted C3-C8cycloalkyl-,halogen, fluoroC1-C6alkyl- wherein the alkyl moiety can be partially orfully fluorinated, cyano, hydroxyl, C1-C6alkoxy, fluoroC1-C6alkoxy-wherein the alkyl moiety can be partially or fully fluorinated, —N(R3)₂,—N(R4)₂, —N(R3)C(O)R3, —C(O)N(R3)₂ and nitro and wherein two R4 moietiesindependently and individually taken from the group consisting ofC1-C6alkyl, branched C3-C6alkyl, hydroxyalkyl-, and alkoxyalkyl andattached to the same nitrogen heteroatom may cyclize to form a C3-C7heterocyclyl ring; k is 0 or 1; n is 0-6; p is 1-4; q is 2-6; r is 0 or1; t is 1-3; v is 1 or 2; m is 0-2; and stereo-, regioisomers andtautomers of such compounds.
 2. Compounds of claim 1 wherein

is selected from the group consisting of

wherein the symbol (**) indicates the point of attachment to thepyridine ring.
 3. Compounds of claim 2 having the formula Ib

wherein A is any possible isomer of pyrazole.
 4. Compounds of claim 3having formula Ic


5. Compounds of claim 3 having formula Id


6. Compounds of claim 3 having formula Ie


7. Compounds of claim 2 having the formula If


8. Compounds of claim 7 having formula Ig


9. Compounds of claim 2 having the formula Ih

wherein A is selected from the group consisting of any possible isomerof phenyl and pyridine.
 10. Compounds of claim 9 having formula Ii


11. Compounds of claim 9 having formula Ij


12. Compounds of claim 2 having the formula Ik


13. Compounds of claim 12 having formula Il


14. A method of treating mammalian disease wherein the disease etiologyor progression is at least partially mediated by the kinase activity ofc-Abl kinase, bcr-Abl kinase, Flt-3 kinase, VEGFR-2 kinase mutants,c-Met, PDGFR-alpha kinase, PDGFR-beta kinase, HER-1, HER-2, HER-3,HER-4, FGFR, c-Kit, oncogenic forms thereof, aberrant fusion proteinsthereof and polymorphs of any of the foregoing, comprising the step ofadministering to the mammal a compound of claim
 1. 15. A method of claim14 wherein said kinase is selected from the group consisting of bcr-Ablfusion protein kinases p210, bcr-Abl fusion protein kinases p190,bcr-Abl fusion protein kinases bearing the T315I gatekeeper mutant inthe Abl kinase domain of p210, bcr-Abl fusion protein kinases bearingthe T3315I gatekeeper mutant in the Abl kinase domain of p190, and otherbcr-Abl polymorphs of any of the foregoing kinases.
 16. The method ofclaim 15, wherein said bcr-Abl fusion protein kinases p210 having SEQ IDNO:3 & SEQ ID NO:4, wherein said bcr-Abl fusion protein kinase p190 hasSEQ ID NO:5, wherein said bcr-Abl fusion protein kinases p210 bearingthe T315I mutation in the Abl kinase domain has SEQ ID NO:6 & SEQ IDNO:7, and wherein said bcr-Abl fusion protein kinase p190 bearing theT315I mutation in the Abl kinase domain has SEQ ID NO:8.
 17. A method ofclaim 14 wherein said kinase is selected from the group consisting ofckit protein kinase, PDGFR-alpha kinase, and any fusion protein,mutation and polymorphs of any of the foregoing.
 18. A method of claim14 wherein said kinase is selected from the group consisting of c-Metprotein kinase, and any fusion protein, mutation and polymorphs of anyof the foregoing.
 19. A pharmaceutical composition comprising a compoundof claim 1, together with a pharmaceutically acceptable carrier,optionally containing an additive selected from the group includingadjuvants, excipients, diluents, and stabilizers.
 20. A method oftreating an individual suffering from a condition selected from thegroup consisting of cancer, hyperproliferative diseases, metabolicdiseases, neurodegenerative diseases, or diseases characterized byangiogenesis, such as solid tumors, melanomas, glioblastomas, ovariancancer, pancreatic cancer, prostate cancer, lung cancers, breastcancers, renal cancers, hepatic cancers, cervical carcinomas, metastasisof primary tumor sites, myeloproliferative diseases, chronic myelogenousleukemia, leukemias, papillary thyroid carcinoma, non-small cell lungcancer, mesothelioma, hypereosinophilic syndrome, gastrointestinalstromal tumors, colonic cancers, ocular diseases characterized byhyperproliferation leading to blindness including retinopathies,diabetic retinopathy, age-related macular degeneration andhypereosinophilic syndrome, rheumatoid arthritis, asthma, chronicobstructive pulmonary, mastocytosis, mast cell leukemia, or disease adisease caused by c-Kit kinase, oncogenic forms thereof, aberrant fusionproteins thereof and polymorphs thereof, comprising the step ofadministering to such individual a compound of claim
 1. 21. The methodof claim 20, said compound being administered by a method selected fromthe group consisting of oral, parenteral, inhalation, and subcutaneous.